Ectonucleotidase inhibitors and methods of use thereof

ABSTRACT

The invention relates to novel heterocyclic compounds and pharmaceutical preparations thereof. The invention further relates to methods of treating or preventing cancer using the novel heterocyclic compounds of the invention.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No. 16/720,369 filed Dec. 19, 2019, which is a divisional of U.S. application Ser. No. 15/851,166 filed Dec. 21, 2017, which claims the benefit of priority to U.S. Provisional Patent Application Nos. 62/437,915 filed Dec. 22, 2016, 62/437,935 filed Dec. 22, 2016 and 62/555,791 filed Sep. 8, 2017, each of which are hereby incorporated by reference in its entirety.

BACKGROUND

CD73, also referred to as 5′-nucleotidase (5′-NT) or ecto-5′-nucleotidase (Ecto 5′NTase), is a membrane-bound cell surface enzyme whose primary role is to catalyze the conversion of extracellular nucleotides (e.g., AMP) to their corresponding nucleosides (e.g., adenosine). CD73 is found in most tissues and expressed on lymphocytes, endothelial cells, and epithelial cells. It is also widely expressed in many tumor cell lines and, notably, is upregulated in cancerous tissues (Antonioli et al., Nat. Rev. Cancer, 13: 842-857, 2013).

In tandem with CD39 (ecto-ATPase), CD73 generates adenosine from ATP/AMP, which is often released from damaged or inflamed cells into the extracellular environment. Extracellular adenosine produced by CD73 interacts with G-protein coupled receptors on target cells. An important downstream effect of this signaling is increased immunosuppression via a number of pathways. For example, CD73 is a co-signaling molecule on T lymphocytes. Under normal circumstances, extracellular adenosine levels promote a self-limiting immune response that prevents excessive inflammation and tissue damage. For tumors, an advantage of abnormally increased CD73 is that the resulting increased CD73-catalyzed adenosine levels yield inhibition of anti-tumor immune system responses.

Even though CD73 plays a role in cancer immunosuppression, higher expression of CD73 is associated with a variety of stages of tumor progression, including tumor vascularization, invasiveness, and metastasis, and with shorter breast cancer patient survival time. Some of these observations result from CD73's enzyme-independent function as an adhesion molecule required for lymphocyte binding to the endothelium.

Overall, CD73 has become an important target for developing new cancer therapies, either as single agents or in combination with other cancer therapies. Indeed, combining CD73 monoclonal antibodies with antibodies for other chemotherapy targets enhances response and survival in animal cancer models (Allard et al., Clin. Cancer Res., 19:5626-35, 2013).

Many of the current cancer treatments and chemotherapeutic agents fail to successfully treat all patients or all symptoms in treated patients, and many of these therapies are associated with undesirable side effects. As certain cancers develop resistance to various chemotherapeutic agents, alternate cancer therapies are needed. Thus, there is a need for additional compounds and methods for treating cancer and other diseases.

SUMMARY

Disclosed herein are compounds of Formula (I):

or a pharmaceutically acceptable salt and/or prodrug thereof, wherein

X is O, NR⁷ or CR⁷R⁸;

Y is O or S;

Z is NR¹⁹, O or S;

Het is heterocyclyl or heteroaryl;

R^(1a) is selected from H, halo, hydroxy, cyano, azido, amino, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, amino-C₁₋₆alkyl, C₁₋₆acyloxy, —O—C(O)—O—C₁₋₆alkyl, C₁₋₆alkoxy, C₂₋₆alkenyl, and C₂₋₆alkynyl; and

R^(1b) is selected from H, halo, C₁₋₆alkyl, hydroxy-C₁₋₆alkyl, amino-C₁₋₆alkyl, C₂₋₆alkenyl, and C₂₋₆alkynyl; or

R^(1a) and R^(1b), together with the carbon atom to which they are attached, form a C═CH₂ or C═C(H)C₁₋₆alkyl;

R^(2a) is selected from H, halo, hydroxy, cyano, azido, amino, C₁₋₆alkyl, hydroxy-C₁₋₆alkyl, amino-C₁₋₆alkyl, C₁₋₆acyloxy, —O—C(O)—O—C₁₋₆alkyl, C₁₋₆alkoxy, C₂₋₆alkenyl, and C₂₋₆alkynyl;

R^(2b) is selected from H, halo, C₁₋₆alkyl, hydroxy-C₁₋₆alkyl, amino-C₁₋₆alkyl, C₂₋₆alkenyl, and C₂₋₆alkynyl; or R¹ and R², together with the carbon atom to which they are attached, form a C═CH₂ or C═C(H)C₁₋₆alkyl;

R³ is selected from H, alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and —(CH₂)—C(O)OR₉;

R⁴ is selected from heteroaryl, alkyl, —C(O)OR⁹, —C(O)NR¹¹R¹², —S(O)₂R¹⁰, —P(O)(OR¹¹)(OR¹²), and —P(O)(OR¹)(NR¹³R¹⁵);

R⁵ is selected from H, cyano, alkyl, cycloalkylalkyl, heterocyclylalkyl, aralkyl, heteroaralkyl, and —C(O)OR⁹;

R₆ is selected from —C(O)OR⁹ and —P(O)(OR¹¹)(OR¹²);

each R⁷ and R⁸ is H

R⁹ is independently selected from H, alkyl, acyloxyalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl, and —(CHR¹³)_(m)—Z—C(O)—R¹⁴;

each R¹⁰ is independently selected from alkyl, alkenyl, alkynyl, amino, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl; and each R¹¹ and R¹² is independently selected from H, alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and —(CHR¹³)_(m)—Z—C(O)—R¹⁴; or

R¹¹ and R¹², together with the atoms to which they are attached, form a 5- to 7-membered heterocyclyl; and

each R¹³ is independently H or alkyl;

each R¹⁴ is independently selected from alkyl, aminoalkyl, heterocyclyl, and heterocyclylalkyl;

R¹⁵ is selected from alkyl, aralkyl, —C(R¹⁶)(R¹⁷)—C(O)O—R¹⁸;

each R¹⁶ and R¹⁷ are selected from H, alkyl, amino-alkyl, hydroxy-alkyl, mercapto-alkyl, sulfonyl-alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, heteroaralkyl, and —(CH₂)C(O)OR⁹;

R¹⁸ is selected from H, alkyl, alkoxyalkyl, aminoalkyl, haloalkyl, amido, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl;

R¹⁹ is H or alkyl, preferably H; and

m is 1 or 2;

provided that either R⁴ is tetrazolyl, or R⁵ is aralkyl or heteroaralkyl, or both.

In certain embodiments, the present invention provides a pharmaceutical composition suitable for use in a subject in the treatment or prevention of cancer comprising an effective amount of any of the compounds described herein (e.g., a compound of the invention, such as a compound of Formula (I), or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical preparations may be for use in treating or preventing a condition or disease as described herein.

Disclosed herein are methods of treating diseases and conditions that benefit from the inhibition of CD73, comprising administering to a subject in need thereof an effective amount of a compound as disclosed herein (e.g., a compound of Formula (I) or any of the embodiments thereof disclosed herein). In certain embodiments, the human subject is in need of such treatment. These diseases include, but are not limited to cancers, such as lung cancer, kidney cancer, skin cancer, breast cancer, and ovarian cancer. Other diseases and conditions that can be treated using the methods described herein include, but are not limited to, neurological, neurodegenerative and CNS disorders and diseases such as depression and Parkinson's disease, cerebral and cardiac ischemic diseases, sleep disorders, fibrosis, immune and inflammatory disorders.

Provided herein are combination therapies of compounds of formula (I) with monoclonal antibodies and other chemotherapeutic agents that can enhance the therapeutic benefit beyond the ability of the adjuvant therapy alone.

DETAILED DESCRIPTION Definitions

Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art of the present disclosure. The following references provide one of skill with a general definition of many of the terms used in this disclosure: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them below, unless specified otherwise.

In some embodiments, chemical structures are disclosed with a corresponding chemical name. In case of conflict, the chemical structure controls the meaning, rather than the name.

In this disclosure, “comprises,” “comprising,” “containing” and “having” and the like can have the meaning ascribed to them in U.S. Patent law and can mean “includes,” “including,” and the like; “consisting essentially of” or “consists essentially” likewise has the meaning ascribed in U.S. Patent law and the term is open-ended, allowing for the presence of more than that which is recited so long as basic or novel characteristics of that which is recited is not changed by the presence of more than that which is recited, but excludes prior art embodiments.

Unless specifically stated or obvious from context, as used herein, the term “or” is understood to be inclusive. Unless specifically stated or obvious from context, as used herein, the terms “a”, “an”, and “the” are understood to be singular or plural.

The term “acyl” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)—, preferably alkylC(O)—.

The term “acylamino” is art-recognized and refers to an amino group substituted with an acyl group and may be represented, for example, by the formula hydrocarbylC(O)NH—.

The term “acyloxy” is art-recognized and refers to a group represented by the general formula hydrocarbylC(O)O—, preferably alkylC(O)O—.

The term “alkoxy” refers to an alkyl group, preferably a lower alkyl group, having an oxygen attached thereto. Representative alkoxy groups include methoxy, ethoxy, propoxy, tert-butoxy and the like.

The term “alkoxyalkyl” refers to an alkyl group substituted with an alkoxy group and may be represented by the general formula alkyl-O-alkyl.

The term “alkenyl”, as used herein, refers to an aliphatic group containing at least one double bond and is intended to include both “unsubstituted alkenyls” and “substituted alkenyls”, the latter of which refers to alkenyl moieties having substituents replacing a hydrogen on one or more carbons of the alkenyl group. Such substituents may occur on one or more carbons that are included or not included in one or more double bonds. Moreover, such substituents include all those contemplated for alkyl groups, as discussed below, except where stability is prohibitive. For example, substitution of alkenyl groups by one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

An “alkyl” group or “alkane” is a straight chained or branched non-aromatic hydrocarbon which is completely saturated. Typically, a straight chained or branched alkyl group has from 1 to about 20 carbon atoms, preferably from 1 to about 10 unless otherwise defined. Examples of straight chained and branched alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl and octyl. A C₁-C₆ straight chained or branched alkyl group is also referred to as a “lower alkyl” group.

Moreover, the term “alkyl” (or “lower alkyl”) as used throughout the specification, examples, and claims is intended to include both “unsubstituted alkyls” and “substituted alkyls”, the latter of which refers to alkyl moieties having substituents replacing a hydrogen on one or more carbons of the hydrocarbon backbone. Such substituents, if not otherwise specified, can include, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxy, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that the moieties substituted on the hydrocarbon chain can themselves be substituted, if appropriate.

For instance, the substituents of a substituted alkyl may include substituted and unsubstituted forms of amino, azido, imino, amido, phosphoryl (including phosphonate and phosphinate), sulfonyl (including sulfate, sulfonamido, sulfamoyl and sulfonate), and silyl groups, as well as ethers, alkylthios, carbonyls (including ketones, aldehydes, carboxylates, and esters), —CF₃, —CN and the like. Exemplary substituted alkyls are described below. Cycloalkyls can be further substituted with alkyls, alkenyls, alkoxys, alkylthios, aminoalkyls, carbonyl-substituted alkyls, —CF₃, —CN, and the like.

The term “C_(x-y)” when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups that contain from x to y carbons in the chain. For example, the term “C_(x-y)alkyl” refers to substituted or unsubstituted saturated hydrocarbon groups, including straight-chain alkyl and branched-chain alkyl groups that contain from x to y carbons in the chain, including haloalkyl groups such as trifluoromethyl and 2,2,2-trifluoroethyl, etc. C₀ alkyl indicates a hydrogen where the group is in a terminal position, a bond if internal. The terms “C_(2-y)alkenyl” and “C_(2-y)alkynyl” refer to substituted or unsubstituted unsaturated aliphatic groups analogous in length and possible substitution to the alkyls described above, but that contain at least one double or triple bond respectively.

The term “alkylamino”, as used herein, refers to an amino group substituted with at least one alkyl group.

The term “alkylthio”, as used herein, refers to a thiol group substituted with an alkyl group and may be represented by the general formula alkylS—.

The term “alkynyl”, as used herein, refers to an aliphatic group containing at least one triple bond and is intended to include both “unsubstituted alkynyls” and “substituted alkynyls”, the latter of which refers to alkynyl moieties having substituents replacing a hydrogen on one or more carbons of the alkynyl group. Such substituents may occur on one or more carbons that are included or not included in one or more triple bonds. Moreover, such substituents include all those contemplated for alkyl groups, as discussed above, except where stability is prohibitive. For example, substitution of alkynyl groups by one or more alkyl, carbocyclyl, aryl, heterocyclyl, or heteroaryl groups is contemplated.

The term “amide”, as used herein, refers to a group

wherein each R³⁰ independently represents a hydrogen or hydrocarbyl group, or two R³⁰ are taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure.

The terms “amine” and “amino” are art-recognized and refer to both unsubstituted and substituted amines and salts thereof, e.g., a moiety that can be represented by

wherein each R³¹ independently represents a hydrogen or a hydrocarbyl group, or two R³¹ are taken together with the N atom to which they are attached complete a heterocycle having from 4 to 8 atoms in the ring structure. The term “aminoalkyl”, as used herein, refers to an alkyl group substituted with an amino group.

The term “aralkyl”, as used herein, refers to an alkyl group substituted with an aryl group.

The term “aryl” as used herein include substituted or unsubstituted single-ring aromatic groups in which each atom of the ring is carbon. Preferably, the ring is a 5- to 7-membered ring, more preferably a 6-membered ring. The term “aryl” also includes polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is aromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Aryl groups include benzene, naphthalene, phenanthrene, phenol, aniline, and the like.

The term “carbamate” is art-recognized and refers to a group

wherein R³² and R³³ independently represent hydrogen or a hydrocarbyl group, such as an alkyl group, or R³² and R³³ taken together with the intervening atom(s) complete a heterocycle having from 4 to 8 atoms in the ring structure.

The terms “carbocycle”, and “carbocyclic”, as used herein, refers to a saturated or unsaturated ring in which each atom of the ring is carbon. The term carbocycle includes both aromatic carbocycles and non-aromatic carbocycles. Non-aromatic carbocycles include both cycloalkane rings, in which all carbon atoms are saturated, and cycloalkene rings, which contain at least one double bond.

The term “carbocycle” includes 5-7 membered monocyclic and 8-12 membered bicyclic rings. Each ring of a bicyclic carbocycle may be selected from saturated, unsaturated and aromatic rings. Carbocycle includes bicyclic molecules in which one, two or three or more atoms are shared between the two rings. The term “fused carbocycle” refers to a bicyclic carbocycle in which each of the rings shares two adjacent atoms with the other ring. Each ring of a fused carbocycle may be selected from saturated, unsaturated and aromatic rings. In an exemplary embodiment, an aromatic ring, e.g., phenyl, may be fused to a saturated or unsaturated ring, e.g., cyclohexane, cyclopentane, or cyclohexene. Any combination of saturated, unsaturated and aromatic bicyclic rings, as valence permits, is included in the definition of carbocyclic. Exemplary “carbocycles” include cyclopentane, cyclohexane, bicyclo[2.2.1]heptane, 1,5-cyclooctadiene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]oct-3-ene, naphthalene and adamantane. Exemplary fused carbocycles include decalin, naphthalene, 1,2,3,4-tetrahydronaphthalene, bicyclo[4.2.0]octane, 4,5,6,7-tetrahydro-1H-indene and bicyclo[4.1.0]hept-3-ene. “Carbocycles” may be substituted at any one or more positions capable of bearing a hydrogen atom.

A “cycloalkyl” group is a cyclic hydrocarbon which is completely saturated. “Cycloalkyl” includes monocyclic and bicyclic rings. Typically, a monocyclic cycloalkyl group has from 3 to about 10 carbon atoms, more typically 3 to 8 carbon atoms unless otherwise defined. The second ring of a bicyclic cycloalkyl may be selected from saturated, unsaturated and aromatic rings. Cycloalkyl includes bicyclic molecules in which one, two or three or more atoms are shared between the two rings. The term “fused cycloalkyl” refers to a bicyclic cycloalkyl in which each of the rings shares two adjacent atoms with the other ring. The second ring of a fused bicyclic cycloalkyl may be selected from saturated, unsaturated and aromatic rings. A “cycloalkenyl” group is a cyclic hydrocarbon containing one or more double bonds.

The term “carbocyclylalkyl”, as used herein, refers to an alkyl group substituted with a carbocycle group.

The term “carbonate” is art-recognized and refers to a group —OCO₂—R³⁴, wherein R³⁴ represents a hydrocarbyl group.

The term “carboxy”, as used herein, refers to a group represented by the formula —CO₂H.

The term “ester”, as used herein, refers to a group —C(O)OR³⁵ wherein R³⁵ represents a hydrocarbyl group.

The term “ether”, as used herein, refers to a hydrocarbyl group linked through an oxygen to another hydrocarbyl group. Accordingly, an ether substituent of a hydrocarbyl group may be hydrocarbyl-O—. Ethers may be either symmetrical or unsymmetrical.

Examples of ethers include, but are not limited to, heterocycle-O-heterocycle and aryl-O-heterocycle. Ethers include “alkoxyalkyl” groups, which may be represented by the general formula alkyl-O-alkyl.

The terms “halo” and “halogen” as used herein means halogen and includes chloro, fluoro, bromo, and iodo.

The terms “hetaralkyl” and “heteroaralkyl”, as used herein, refers to an alkyl group substituted with a hetaryl group.

The term “heteroalkyl”, as used herein, refers to a saturated or unsaturated chain of carbon atoms and at least one heteroatom, wherein no two heteroatoms are adjacent.

The terms “heteroaryl” and “hetaryl” include substituted or unsubstituted aromatic single ring structures, preferably 5- to 7-membered rings, more preferably 5- to 6-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heteroaryl” and “hetaryl” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heteroaromatic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heteroaryl groups include, for example, pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrazine, pyridazine, and pyrimidine, and the like.

The term “heteroatom” as used herein means an atom of any element other than carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, and sulfur.

The terms “heterocyclyl”, “heterocycle”, and “heterocyclic” refer to substituted or unsubstituted non-aromatic ring structures, preferably 3- to 10-membered rings, more preferably 3- to 7-membered rings, whose ring structures include at least one heteroatom, preferably one to four heteroatoms, more preferably one or two heteroatoms. The terms “heterocyclyl” and “heterocyclic” also include polycyclic ring systems having two or more cyclic rings in which two or more carbons are common to two adjoining rings wherein at least one of the rings is heterocyclic, e.g., the other cyclic rings can be cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls. Heterocyclyl groups include, for example, piperidine, piperazine, pyrrolidine, morpholine, lactones, lactams, and the like.

The term “heterocyclylalkyl”, as used herein, refers to an alkyl group substituted with a heterocycle group.

The term “hydrocarbyl”, as used herein, refers to a group that is bonded through a carbon atom that does not have a ═O or ═S substituent, and typically has at least one carbon-hydrogen bond and a primarily carbon backbone, but may optionally include heteroatoms. Thus, groups like methyl, ethoxyethyl, 2-pyridyl, and trifluoromethyl are considered to be hydrocarbyl for the purposes of this application, but substituents such as acetyl (which has a ═O substituent on the linking carbon) and ethoxy (which is linked through oxygen, not carbon) are not. Hydrocarbyl groups include, but are not limited to aryl, heteroaryl, carbocycle, heterocyclyl, alkyl, alkenyl, alkynyl, and combinations thereof.

The term “hydroxyalkyl”, as used herein, refers to an alkyl group substituted with a hydroxy group.

The term “lower” when used in conjunction with a chemical moiety, such as, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy is meant to include groups where there are ten or fewer non-hydrogen atoms in the substituent, preferably six or fewer. A “lower alkyl”, for example, refers to an alkyl group that contains ten or fewer carbon atoms, preferably six or fewer. In certain embodiments, acyl, acyloxy, alkyl, alkenyl, alkynyl, or alkoxy substituents defined herein are respectively lower acyl, lower acyloxy, lower alkyl, lower alkenyl, lower alkynyl, or lower alkoxy, whether they appear alone or in combination with other substituents, such as in the recitations hydroxyalkyl and aralkyl (in which case, for example, the atoms within the aryl group are not counted when counting the carbon atoms in the alkyl substituent).

The terms “polycyclyl”, “polycycle”, and “polycyclic” refer to two or more rings (e.g., cycloalkyls, cycloalkenyls, cycloalkynyls, aryls, heteroaryls, and/or heterocyclyls) in which two or more atoms are common to two adjoining rings, e.g., the rings are “fused rings”. Each of the rings of the polycycle can be substituted or unsubstituted. In certain embodiments, each ring of the polycycle contains from 3 to 10 atoms in the ring, preferably from 5 to 7.

The term “silyl” refers to a silicon moiety with three hydrocarbyl moieties attached thereto.

The term “substituted” refers to moieties having substituents replacing a hydrogen on one or more carbons of the backbone. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom and the substituent, and that the substitution results in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. As used herein, the term “substituted” is contemplated to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, aromatic and non-aromatic substituents of organic compounds. The permissible substituents can be one or more and the same or different for appropriate organic compounds. For purposes of this invention, the heteroatoms such as nitrogen may have hydrogen substituents and/or any permissible substituents of organic compounds described herein which satisfy the valences of the heteroatoms. Substituents can include any substituents described herein, for example, a halogen, a hydroxyl, a carbonyl (such as a carboxyl, an alkoxycarbonyl, a formyl, or an acyl), a thiocarbonyl (such as a thioester, a thioacetate, or a thioformate), an alkoxy, a phosphoryl, a phosphate, a phosphonate, a phosphinate, an amino, an amido, an amidine, an imine, a cyano, a nitro, an azido, a sulfhydryl, an alkylthio, a sulfate, a sulfonate, a sulfamoyl, a sulfonamido, a sulfonyl, a heterocyclyl, an aralkyl, or an aromatic or heteroaromatic moiety. It will be understood by those skilled in the art that substituents can themselves be substituted, if appropriate. Unless specifically stated as “unsubstituted,” references to chemical moieties herein are understood to include substituted variants. For example, reference to an “aryl” group or moiety implicitly includes both substituted and unsubstituted variants.

The term “sulfate” is art-recognized and refers to the group —OSO₃H, or a pharmaceutically acceptable salt thereof.

The term “sulfonamide” is art-recognized and refers to the group represented by the general formulae

wherein R³⁶ and R³⁷ independently represent hydrogen or hydrocarbyl, such as alkyl, or R³⁶ and R³⁷ taken together with the intervening atom(s) complete a heterocycle having from 4 to 8 atoms in the ring structure.

The term “sulfoxide” is art-recognized and refers to the group —S(O)—R³⁸, wherein R³⁸ represents a hydrocarbyl.

The term “sulfonate” is art-recognized and refers to the group SO₃H, or a pharmaceutically acceptable salt thereof.

The term “sulfone” is art-recognized and refers to the group —S(O)₂—R³⁹, wherein R³⁹ represents a hydrocarbyl.

The term “thioalkyl”, as used herein, refers to an alkyl group substituted with a thiol group.

The term “thioester”, as used herein, refers to a group —C(O)SR⁴⁰ or —SC(O)R⁴⁰ wherein R¹⁰ represents a hydrocarbyl.

The term “thioether”, as used herein, is equivalent to an ether, wherein the oxygen is replaced with a sulfur.

The term “urea” is art-recognized and may be represented by the general formula

wherein R⁴¹ and R⁴² independently represent hydrogen or a hydrocarbyl, such as alkyl, or either occurrence of R⁴¹ taken together with R⁴² and the intervening atom(s)complete a heterocycle having from 4 to 8 atoms in the ring structure.

The term “protecting group” refers to a group of atoms that, when attached to a reactive functional group in a molecule, mask, reduce or prevent the reactivity of the functional group. Typically, a protecting group may be selectively removed as desired during the course of a synthesis. Examples of protecting groups can be found in Greene and Wuts, Protective Groups in Organic Chemistry, 3^(rd) Ed., 1999, John Wiley & Sons, NY and Harrison et al., Compendium of Synthetic Organic Methods, Vols. 1-8, 1971-1996, John Wiley & Sons, NY. Representative nitrogen protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (“CBZ”), tert-butoxycarbonyl (“Boc”), trimethylsilyl (“TMS”), 2-trimethylsilyl-ethanesulfonyl (“TES”), trityl and substituted trityl groups, allyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (“FMOC”), nitro-veratryloxycarbonyl (“NVOC”) and the like. Representative hydroxyl protecting groups include, but are not limited to, those where the hydroxyl group is either acylated (esterified) or alkylated such as benzyl and trityl ethers, as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers (e.g., TMS or TIPS groups), glycol ethers, such as ethylene glycol and propylene glycol derivatives and allyl ethers.

In the pictorial representation of the compounds given through this application, a thickened tapered line (

) indicates a substituent which is above the plane of the ring to which the asymmetric carbon belongs and a dotted line (

) indicates a substituent which is below the plane of the ring to which the asymmetric carbon belongs.

In certain embodiments, compounds of the invention may be racemic. In certain embodiments, compounds of the invention may be enriched in one enantiomer. For example, a compound of the invention may have greater than about 30% ee, about 40% ee, about 50% ee, about 60% ee, about 70% ee, about 80% ee, about 90% ee, or even about 95% or greater ee. In certain embodiments, compounds of the invention may have more than one stereocenter. In certain such embodiments, compounds of the invention may be enriched in one or more diastereomer. For example, a compound of the invention may have greater than about 30% de, about 40% de, about 50% de, about 60% de, about 70% de, about 80% de, about 90% de, or even about 95% or greater de.

In certain embodiments, the therapeutic preparation may be enriched to provide predominantly one enantiomer of a compound (e.g., of Formula (I)). An enantiomerically enriched mixture may comprise, for example, at least about 60 mol percent of one enantiomer, or more preferably at least about 75, about 90, about 95, or even about 99 mol percent. In certain embodiments, the compound enriched in one enantiomer is substantially free of the other enantiomer, wherein substantially free means that the substance in question makes up less than about 10%, or less than about 5%, or less than about 4%, or less than about 3%, or less than about 2%, or less than about 1% as compared to the amount of the other enantiomer, e.g., in the composition or compound mixture. For example, if a composition or compound mixture contains about 98 grams of a first enantiomer and about 2 grams of a second enantiomer, it would be said to contain about 98 mol percent of the first enantiomer and only about 2% of the second enantiomer.

In certain embodiments, the therapeutic preparation may be enriched to provide predominantly one diastereomer of a compound (e.g., of Formula (I)). A diastereomerically enriched mixture may comprise, for example, at least about 60 mol percent of one diastereomer, or more preferably at least about 75, about 90, about 95, or even about 99 mol percent.

The term “subject” to which administration is contemplated includes, but is not limited to, humans (i.e., a male or female of any age group, e.g., a pediatric subject (e.g., infant, child, adolescent) or adult subject (e.g., young adult, middle-aged adult or senior adult)) and/or other primates (e.g., cynomolgus monkeys, rhesus monkeys); mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, goats, cats, and/or dogs; and/or birds, including commercially relevant birds such as chickens, ducks, geese, quail, and/or turkeys. Preferred subjects are humans.

As used herein, a therapeutic that “prevents” a disorder or condition refers to a compound that, in a statistical sample, reduces the occurrence of the disorder or condition in the treated sample relative to an untreated control sample, or delays the onset or reduces the severity of one or more symptoms of the disorder or condition relative to the untreated control sample.

The term “treating” includes prophylactic and/or therapeutic treatments. The term “prophylactic or therapeutic” treatment is art-recognized and includes administration to the subject of one or more of the disclosed compositions. If it is administered prior to clinical manifestation of the unwanted condition (e.g., disease or other unwanted state of the subject) then the treatment is prophylactic (i.e., it protects the subject against developing the unwanted condition), whereas if it is administered after manifestation of the unwanted condition, the treatment is therapeutic, (i.e., it is intended to diminish, ameliorate, or stabilize the existing unwanted condition or side effects thereof).

The term “prodrug” is intended to encompass compounds which, under physiologic conditions, are converted into the therapeutically active agents of the present invention (e.g., a compound of Formula (I)). A common method for making a prodrug is to include one or more selected moieties which are hydrolyzed under physiologic conditions to reveal the desired molecule. In other embodiments, the prodrug is converted by an enzymatic activity of the subject. For example, esters or carbonates (e.g., esters or carbonates of alcohols or carboxylic acids) are preferred prodrugs of the present invention. In certain embodiments, some or all of the compounds of Formula (I) in a formulation represented above can be replaced with the corresponding suitable prodrug, e.g., wherein a hydroxyl in the parent compound is presented as an ester or a carbonate or carboxylic acid.

An “effective amount”, as used herein, refers to an amount that is sufficient to achieve a desired biological effect. A “therapeutically effective amount”, as used herein refers to an amount that is sufficient to achieve a desired therapeutic effect. For example, a therapeutically effective amount can refer to an amount that is sufficient to improve at least one sign or symptom of cancer.

A “response” to a method of treatment can include a decrease in or amelioration of negative symptoms, a decrease in the progression of a disease or symptoms thereof, an increase in beneficial symptoms or clinical outcomes, a lessening of side effects, stabilization of disease, partial or complete remedy of disease, among others.

The present invention provides compounds of Formula (I):

or a pharmaceutically acceptable salt and/or prodrug thereof, wherein

X is O, NR⁷ or CR⁷R⁸;

Y is O or S;

Z is NR¹⁹, O or S;

Het is heterocyclyl or heteroaryl;

R^(1a) is selected from H, halo, hydroxy, cyano, azido, amino, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, amino-C₁₋₆alkyl, C₁₋₆acyloxy, —O—C(O)—O—C₁₋₆alkyl, C₁₋₆alkoxy, C₂₋₆alkenyl, and C₂₋₆alkynyl; and

R^(1b) is selected from H, halo, C₁₋₆alkyl, hydroxy-C₁₋₆alkyl, amino-C₁₋₆alkyl, C₂₋₆alkenyl, and C₂₋₆alkynyl; or

R^(1a) and R^(1b), together with the carbon atom to which they are attached, form a C═CH₂ or C═C(H)C₁₋₆alkyl;

R^(2a) is selected from H, halo, hydroxy, cyano, azido, amino, C₁₋₆alkyl, hydroxy-C₁₋₆alkyl, amino-C₁₋₆alkyl, C₁₋₆acyloxy, —O—C(O)—O—C₁₋₆alkyl, C₁₋₆alkoxy, C₂₋₆alkenyl, and C₂₋₆alkynyl;

R^(2b) is selected from H, halo, C₁₋₆alkyl, hydroxy-C₁₋₆alkyl, amino-C₁₋₆alkyl, C₂₋₆alkenyl, and C₂₋₆alkynyl; or

R^(2a) and R^(2b), together with the carbon atom to which they are attached, form a C═CH₂ or C═C(H)C₁₋₆alkyl;

R³ is selected from H, alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and —(CH₂)—C(O)OR₉;

R⁴ is selected from heteroaryl, alkyl, —C(O)OR⁹, —C(O)NR¹¹R¹², —S(O)₂R¹⁰, —P(O)(OR¹¹)(OR¹²), and —P(O)(OR¹¹)(NR¹³R¹⁵);

R⁵ is selected from H, cyano, alkyl, cycloalkylalkyl, heterocyclylalkyl, aralkyl, heteroaralkyl, and —C(O)OR⁹;

R₆ is selected from —C(O)OR⁹ and —P(O)(OR¹¹)(OR¹²);

each R⁷ and R⁸ is H

R⁹ is independently selected from H, alkyl, acyloxyalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl, and —(CHR¹³)_(m)—Z—C(O)—R¹⁴;

each R¹⁰ is independently selected from alkyl, alkenyl, alkynyl, amino, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl; and

each R¹¹ and R¹² is independently selected from H, alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and —(CHR¹³)_(m)—Z—C(O)—R¹⁴; or

R¹¹ and R¹², together with the atoms to which they are attached, form a 5- to 7-membered heterocyclyl; and

each R¹³ is independently H or alkyl;

each R¹⁴ is independently selected from alkyl, aminoalkyl, heterocyclyl, and heterocyclylalkyl;

R¹⁵ is selected from alkyl, aralkyl, —C(R¹⁶)(R¹⁷)—C(O)O—R¹⁸;

each R¹⁶ and R¹⁷ are selected from H, alkyl, amino-alkyl, hydroxy-alkyl, mercapto-alkyl, sulfonyl-alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, heteroaralkyl, and —(CH₂)C(O)OR⁹;

R¹⁸ is selected from H, alkyl, alkoxyalkyl, aminoalkyl, haloalkyl, amido, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl;

R¹⁹ is H or alkyl, preferably H; and

m is 1 or 2;

provided that either R⁴ is tetrazolyl, or R⁵ is aralkyl or heteroaralkyl, or both.

In certain embodiments, R^(1a) is selected from H, halo, hydroxy, cyano, azido, amino, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, amino-C₁₋₆alkyl, C₁₋₆acyloxy, C₁₋₆alkoxy, C₂₋₆alkenyl, and C₂₋₆alkynyl. In certain embodiments, R^(1a) is fluoro, chloro or bromo, preferably fluoro. In certain embodiments, R^(1a) is C₁₋₆alkoxy. In certain embodiments, R^(1a) is C₁₋₆alkyl. In certain embodiments, R^(1a) is hydroxy. In certain embodiments, R^(1a) is ethynyl or vinyl. In certain embodiments, R^(1a) is cyano. In certain embodiments, R^(1a) is azido. In certain embodiments, R^(1a) is amino. In certain embodiments, R^(1a) is hydrogen.

In certain embodiments, R^(2a) is selected from H, halo, hydroxy, cyano, azido, amino, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, amino-C₁₋₆alkyl, C₁₋₆acyloxy, C₁₋₆alkoxy, C₂₋₆alkenyl, and C₂₋₆alkynyl. In other embodiments, R^(2a) is fluoro, chloro, or bromo, preferably fluoro. In certain embodiments, R^(2a) is C₁₋₆alkoxy. In certain embodiments, R^(2a) is C₁₋₆alkyl. In certain embodiments, R^(2a) is hydroxy. In certain embodiments, R^(2a) is cyano. In certain embodiments, R^(2a) is azido. In certain embodiments, R^(2a) is amino. In certain embodiments, R^(2a) is C₁₋₆acyloxy.

In certain embodiments, R^(1b) is H. In other embodiments, R^(1b) is fluoro. In other embodiments, R^(1b) is C₁₋₆alkyl. In certain embodiments, R^(2b) is H. In other embodiments, R^(2b) is fluoro. In other embodiments, R^(2b) is C₁₋₆alkyl.

In certain embodiments, R^(1a) is fluoro and R^(1b) is H. In other embodiments, R^(2a) is fluoro and R^(2b) is H. In certain embodiments, R^(1a) and R^(1b) are each fluoro. In other embodiments, R^(2a) and R^(2b) are each fluoro. In some embodiments, R^(1a) is fluoro and R^(2a) is C₁₋₆alkoxy. In some embodiments, R^(1a) is fluoro and R^(2a) is C₁₋₆alkyl, such as methyl or ethyl. In certain embodiments, R^(2a) is hydroxy and R^(2b) is methyl.

In certain embodiments, R^(1a) is fluoro and R^(2a) is hydroxy. In certain embodiments, R^(1a) is chloro and R^(2a) is hydroxy. In certain embodiments, R^(1a) is hydroxy and R^(2a) is hydrogen. In certain embodiments, R^(1a) is fluoro and R^(2a) is hydrogen. In certain embodiments, R^(1a) is hydroxy and R^(2a) is C₁₋₆alkoxy, such as methoxy. In certain embodiments, R^(1a) is hydroxy and R^(2a) is fluoro. In certain embodiments, R^(1a) is hydroxy and R^(2a) and R^(2b), together with the carbon atom to which they are attached, form an ethynyl. In certain embodiments, R^(1a) is hydroxy and R^(2a) is C₁₋₆alkyl, such as methyl. In certain embodiments, R^(1a) is fluoro, R^(2a) is fluoro and R^(2a) is hydroxy. In certain embodiments, R^(1a) is hydroxy and R^(2a) is hydroxyl and R^(2b) is C₁₋₆alkynyl, such as ethynyl. In certain embodiments, R^(1a) is hydroxy and R^(2a) is C₁₋₆ alkynyl, such as ethynyl. In certain embodiments, R^(1a) is hydroxy and R^(2a) is C₂₋₆alkenyl. In certain embodiments, R^(1a) is hydroxy and R^(2a) is amino. In certain embodiments, R^(1a) is hydroxy and R^(2a) is azido. In certain embodiments, R^(1a) is azido and R^(2a) is hydroxy. In certain embodiments, R^(1a) is C₁₋₆alkyl, such as methyl, and R^(2a) is hydroxy.

In certain embodiments, the compound of Formula (I) has the following structure:

In certain embodiments, R^(1a) is in the α-configuration. In some such embodiments, the compound of Formula (I) has the structure (IA):

In other embodiments, R^(1a) is in the β-configuration. In some such embodiments, the compound of Formula (I) has the structure (IB):

In certain embodiments, R^(2a) is in the α-configuration. In some such embodiments, the compound of Formula (I) has the structure (IC):

In other embodiments, R^(2a) is in the β-configuration. In some embodiments, the compound of Formula (I) has the structure (ID):

In certain embodiments, the compound of Formula (I) has the structure (IE):

In certain embodiments, R³ is alkyl, and the alkyl is unsubstituted or substituted, e.g., with one or more substituents selected from halo, CN, NO₂, azido, hydroxy, alkoxy, alkylthio, thioalkoxy, carbonyl, thiocarbonyl, amidino, imino, amino, amido, alkoxycarbonyl, carbamate, urea, sulfinamido, sulfonamido, sulfinyl, sulfinamido, sulfonyl, phosphoryl, phosphate, phosphonate, and phosphinate. In certain embodiments, the substituents are selected from halo, CN, azido, alkoxy, carbonyl, amino, amido, and alkoxycarbonyl. In certain preferred embodiments, R³ is H. In other preferred embodiments, R³ is unsubstituted C₁₋₆alkyl.

In certain embodiments, R⁴ is heteroaryl. In certain preferred embodiments, R⁴ is tetrazolyl.

In certain embodiments, R⁴ is selected from —C(O)OR⁹, —C(O)NR¹¹R¹², —S(O)₂R¹⁰, and —P(O)(OR¹¹)(OR¹²). In some preferred embodiments, R⁴ is —C(O)OR⁹. In certain embodiments, R⁹ is H or C₁₋₆alkyl. In some embodiments, R⁴ is —C(O)NR¹¹R¹². In certain such embodiments, R¹¹ and R¹² are each alkyl, such as methyl. In alternative such embodiments, R¹¹ is alkyl and R¹² is hydrogen. In other embodiments, R⁴ is alkyl substituted with carboxyl or ester (e.g., alkoxycarbonyl).

In certain embodiments, R⁵ is selected from H, alkyl, cycloalkylalkyl, heterocyclylalkyl, aralkyl, and heteroaralkyl. In certain embodiments, R⁵ is H, aralkyl, or heteroaralkyl. For example, R⁵ can be —CH₂-pyridinyl, —CH₂-thiophenyl, benzyl or β-naphthyl. In certain embodiments, R⁴ is heteroaryl, such as tetrazolyl, R⁵ is unsubstituted or substituted benzyl and R⁶ is —COOH.

In certain embodiments, R₅ is selected from H, alkyl, cycloalkylalkyl, heterocyclylalkyl, aralkyl, and heteroaralkyl. In certain embodiments, R⁵ is selected from alkyl, cycloalkylalkyl, heterocyclylalkyl, aralkyl and heteroaralkyl, and each is unsubstituted or substituted with one or more substituents, e.g., selected from halo, CN, NO₂, azido, hydroxy, alkoxy, alkylthio, thioalkoxy, carbonyl, thiocarbonyl, amidino, imino, amino, amido, alkoxycarbonyl, carbamate, urea, sulfinamido, sulfonamido, sulfinyl, sulfinamido, sulfonyl, phosphoryl, phosphate, phosphonate, phosphinate, cycloalkyl, heteroaryl, heterocyclyl, arylalkyl and heteroarylalkyl. In certain embodiments, the substituents are selected from halo, CN, NO₂, azido, hydroxy, alkoxy, alkylthio, thioalkoxy, carbonyl, thiocarbonyl, amidino, imino, amino, amido, alkoxycarbonyl, carbamate, urea, sulfinamido, sulfonamido, sulfinyl, sulfinamido, sulfonyl, phosphoryl, phosphate, phosphonate, phosphinate, cycloalkyl, heterocyclyl, arylalkyl and heteroarylalkyl. In certain embodiments, the substituents are selected from halo, CN, azido, alkoxy, carbonyl, amino, amido, and alkoxycarbonyl. In other embodiments, the substitutents are selected from chloro, trifluoromethyl, trifluoromethoxy, phenyloxy, dimethylamido, methylsulfonyl, CN, and carboxylic acid. In some preferred embodiments, R₅ is H or aralkyl.

In some embodiments, R⁵ is aralkyl, e.g., wherein the aryl ring is substituted or unsubstituted phenyl or naphthyl. In other embodiments, R⁵ is heteroaralkyl, e.g., wherein the heteroaryl ring is selected from substituted or unsubstituted benzofuranyl, benzothienyl, benzothiazolyl, pyridyl, thienyl, furanyl, pyrazolyl, thiazolyl, oxazolyl, and oxadiazolyl.

In some embodiments, R⁵ is aralkyl or heteroaralkyl, wherein the aryl or heteroaryl ring, respectively, is unsubstituted or substituted with one or more substituents, e.g., selected from halo, CN, OH, alkyl, alkenyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyloxy, alkylsulfonyl, sulfonamido, amido, amino, carboxyl, ester (e.g., lower alkyl ester), heterocyclyl, heteroaryl, aryl, aralkyl, and heteroaralkyl. In certain embodiments, the substituents are selected from halo, CN, OH, alkyl, alkenyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyloxy, alkylsulfonyl, sulfonamido, amido, amino, carboxyl, ester (e.g., lower alkyl ester), heteroaryl, aryl, aralkyl, and heteroaralkyl. In certain such embodiments, the substituents on the aryl or heteroaryl ring of the aralkyl or heteroaralkyl, respectively, are selected from halo, CN, haloalkyl, haloalkoxy, carboxy, ester (e.g., lower alkyl ester), and aryl. In other such embodiments, the substituents on the aryl or heteroaryl ring of the aralkyl or heteroaralkyl, respectively, are selected from tetrazolyl, substituted or unsubstituted phenyl, or substituted or unsubstituted benzyl.

In certain embodiments, R₆ is —C(O)OR₉. In some embodiments, R₉ is H or C₁₋₆alkyl. In other embodiments, R₆ is —P(O)(OR₁₁)(OR₁₂), and R₁₁ and R₁₂ are each H. In certain embodiments, R₇ is H or C₁₋₆alkyl. In certain embodiments, R₉ is H. In certain embodiments, R₉ is H or C₁₋₆alkyl, such as methyl or ethyl. In certain embodiments, R₁₁ and R₁₂ are each H.

In certain embodiments, each R₇, R₈, R₉, R₁₀, R₁₁ and R₁₂ is independently selected from alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl, and each is unsubstituted or substituted, e.g., with one or more selected from halo, CN, NO₂, azido, hydroxy, alkoxy, alkylthio, thioalkoxy, carbonyl, thiocarbonyl, amidino, imino, amino, amido, alkoxycarbonyl, carbamate, urea, sulfinamido, sulfonamido, sulfinyl, sulfinamido, sulfonyl, phosphoryl, phosphate, phosphonate, and phosphinate. In some embodiments, the substituents are selected from halo, CN, azido, alkoxy, carbonyl, amino, amido, and alkoxycarbonyl.

In certain embodiments, X is O. In certain embodiments, Y is O. In certain embodiments, the Y-bearing substituent is in the R-configuration. In other embodiments, the Y-bearing substituent is in the S-configuration. In certain embodiments, Z is O. In other embodiments, Z is NH.

In certain embodiments, Het is a nitrogen-containing heterocyclyl or heteroaryl. In certain embodiments, Het is attached via a nitrogen atom. In other, embodiments, Het is attached via a carbon atom. In some embodiments, Het is a 5- to 8-membered monocyclic or 5- to 10-membered bicyclic heteroaryl and is unsubstituted or substituted, e.g., with one or more substituents selected from halo, CN, NO₂, azido, hydroxy, alkoxy, alkylthio, thioalkoxy, carbonyl, thiocarbonyl, cyclothioalkyl, amidino, imino, amino, amido, alkoxycarbonyl, carbamate, urea, sulfinamido, sulfonamido, sulfinyl, sulfinamido, sulfonyl, phosphoryl, phosphate, phosphonate, and phosphinate. In certain embodiments, the substituents are selected from halo, CN, NO₂, azido, hydroxy, alkoxy, alkylthio, thioalkoxy, carbonyl, thiocarbonyl, amidino, imino, amino, amido, alkoxycarbonyl, carbamate, urea, sulfinamido, sulfonamido, sulfinyl, sulfinamido, sulfonyl, phosphoryl, phosphate, phosphonate, and phosphinate. In certain embodiments, the substituents are selected from halo, CN, azido, alkoxy, carbonyl, amino, amido, and alkoxycarbonyl. In certain preferred embodiments, Het is selected from purinyl, imidazopyrimidinyl, and pyrrolopyrimidinyl. In other embodiments, Het is substituted with one or two substituents independently selected from halo, aralkyl, amino, azido and hydroxy. In some embodiments, Het is substituted with one halo and one amino substituent. In certain preferred embodiments, Het is

In certain embodiments, Het is selected from

In certain embodiments, Het is

In certain embodiments, Het is selected from

In certain embodiments, Het is a group of formula (i) through (xiv) below:

wherein:

R^(u) is hydrogen, halo, cyano, —NH₂, —NHR²⁰, —NHCOR²⁰, —NR²⁰R²¹, —R²⁰, —SR²⁰, —OH, and —OR²⁰;

R^(w) is hydrogen, halo, —NHR²², —NR²²R²³, —R²², —OH, and —OR²²;

R^(v) and R^(x) are independently hydrogen, halo, haloC₁₋₆alkyl, —NH₂, —NHR²⁴, —NR²⁴R²⁵, —R²⁴, —SR²⁴, cyano, —OH, —OR²⁴, —SO₂R²⁴, —C₁₋₆alkyleneNH₂, —C₁₋₆alkyleneNHR²⁴, —C₁₋₆alkyleneNR²⁴R²⁵, —R²⁴, —C₁₋₆alkyleneSR²⁴, —C₁₋₆alkyleneOH, —C₁₋₆alkyleneOR²⁴, —C₁₋₆alkyleneSO₂R²⁴,

R^(s) and R^(t) are independently hydrogen, halo, or C₁₋₆alkyl; and wherein: R²⁰, R²¹, R²², R²³, R²⁴ and R²⁵ are independently optionally substituted C₁₋₆alkyl, —C₂-C₆alkenyl, —C₂-C₆alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylC₁₋₆alkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylC₁₋₆alkyl, optionally substituted aryl, optionally substituted arylC₁₋₆alkyl, optionally substituted heteroaryl, or optionally substituted heteroarylC₁₋₆alkyl; or R²⁰ and R²¹, R²² and R²³, and R²⁴ and R²⁵ together with the nitrogen to which they are attached form an optionally substituted nitrogen-containing heterocyclyl.

In certain embodiments

represents

In certain embodiments,

represents

In certain embodiments,

represents

In certain embodiments,

represents

In certain embodiments,

represents

wherein

each R_(A) is independently selected from halo, CN, OH, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyloxy (e.g., substituted or unsubstituted benzyloxy), alkylsulfonyl, sulfonamido, amido, amino, hydroxycarbonyl, alkoxycarbonyl, heterocyclyl, heteroaryl, aryl, aralkyl, and heteroaralkyl; and

k is 1, 2, or 3.

In certain embodiments, R_(A) is selected from halo, CN, OH, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, aryloxy, aralkyloxy (e.g., substituted or unsubstituted benzyloxy), alkylsulfonyl, sulfonamido, amido, amino, hydroxycarbonyl, alkoxycarbonyl, heteroaryl, aryl, aralkyl, and heteroaralkyl. In certain embodiments, R_(A) is selected from pyridyl, pyrazolyl, pyrrolidinonyl, azapanonyl, morpholinonyl, piperazonyl, tetrahydropyrimidinonyl, phenyl, and benzyloxy.

FURTHER EMBODIMENTS

In the following numbered embodiments, unless otherwise specified:

-   C₁₋₆alkyl is unsubstituted; -   C₁₋₆alkyl of C₁₋₆alkyloxy is unsubstituted; -   haloC₁₋₆alkyl is C₁₋₆alkyl substituted with one to five halo atoms; -   haloC₁₋₆alkyloxy is C₁₋₆alkyloxy substituted with one to five halo     atoms; -   C₂₋₆alkenyl is unsubstituted; -   C₂₋₆alkynyl is unsubstituted; -   C₁₋₆alkylene group is unsubstituted; -   C₂₋₆alkenylene group is unsubstituted; -   optionally substituted C₁₋₆alkyl is optionally substituted with one     or two substituents independently selected from hydroxy,     C₁₋₆alkyloxy, halo, haloC₁₋₆alkyl, haloC₁₋₆alkyloxy, cyano, —NH₂,     —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂, optionally substituted carbocyclyl,     optionally substituted aryl, optionally substituted heteroaryl, and     optionally substituted heterocyclyl; -   carbocyclyl is an unsubstituted saturated or partially unsaturated     mono or bicyclic ring containing 3 to 10 carbon atoms; -   optionally substituted carbocyclyl is optionally substituted with     one, two, or three substituents independently selected from     C₁₋₆alkyl, halo, hydroxy, or C₁₋₆alkoxy; -   aryl is phenyl or naphthyl; -   optionally substituted aryl is optionally substituted with one, two,     or three substituents independently selected from hydroxy,     C₁₋₆alkyloxy, halo, haloC₁₋₆alkyl, haloC₁₋₆alkyloxy, cyano, —NH₂,     —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂, optionally substituted carbocyclyl,     aryl [optionally substituted with one, two, or three substituents     independently selected from hydroxy, C₁₋₆alkyloxy, halo,     haloC₁₋₆alkyl, haloC₁₋₆alkyloxy, cyano, —NH₂, —NH(C₁₋₆alkyl), and     —N(C₁₋₆alkyl)₂], heteroaryl [optionally substituted with one, two,     or three substituents independently selected from hydroxy,     C₁₋₆alkyloxy, halo, haloC₁₋₆alkyl, haloC₁₋₆alkyloxy, cyano, —NH₂,     —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂], and heterocyclyl [optionally     substituted with one, two, or three substituents independently     selected from hydroxy, C₁₋₆alkyloxy, halo, haloC₁₋₆alkyl,     haloC₁₋₆alkyloxy, cyano, —NH₂, —NH(C₁₋₆alkyl), —N(unsubstitued     C₁₋₆alkyl)₂]; -   heteroaryl is an unsubstituted five to ten membered or five to six     membered aromatic ring containing one to four or one to three     heteroatoms independently selected from N, O, and S, the remaining     ring atoms being carbon; -   optionally substituted heteroaryl is a five to ten membered aromatic     ring containing one to four heteroatoms independently selected from     N, O, and S, the remaining ring atoms being carbon, that is     optionally substituted with one, two, or three substituents     independently selected from hydroxy, C₁₋₆alkyloxy, halo,     haloC₁₋₆alkyl, haloC₁₋₆alkyloxy, cyano, —NH₂, —NH(C₁₋₆alkyl),     —N(C₁₋₆alkyl)₂, optionally substituted carbocyclyl, aryl [optionally     substituted with one, two, or three substituents independently     selected from hydroxy, C₁₋₆alkyloxy, halo, haloC₁₋₆alkyl,     haloC₁₋₆alkyloxy, cyano, —NH₂, —NH(C₁₋₆alkyl), and —N(C₁₋₆alkyl)₂],     heteroaryl [optionally substituted with one, two, or three     substituents independently selected from hydroxy, unsubstituted     C₁₋₆alkyloxy, halo, haloC₁₋₆alkyl, haloC₁₋₆alkyloxy, cyano, —NH₂,     —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂], and heterocyclyl [optionally     substituted with one, two, or three substituents independently     selected from hydroxy, C₁₋₆alkyloxy, halo, haloC₁₋₆alkyl,     haloC₁₋₆alkyloxy, cyano, —NH₂, —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂];     heterocyclyl is an unsubstituted monocyclic, saturated or partially     unsaturated 4 to 8 membered ring containing one to three heteroatoms     independently selected from N, O, S, SO, and SO₂ and optionally     contains one or two CO, the remaining atoms in the ring being carbon     that is optionally fused to phenyl or 5 to 6 membered carbocyclyl or     5 or 6 heteroaryl ring; -   nitrogen-containing heterocyclyl is a heterocyclyl ring that has at     least a nitrogen atom; -   optionally substituted heterocyclyl is heterocyclyl that is     optionally substituted with one, two, or three substituents     independently selected from hydroxy, C₁₋₆alkyloxy, halo,     haloC₁₋₆alkyl, haloC₁₋₆alkyloxy, cyano, —NH₂, —NH(C₁₋₆alkyl),     —N(C₁₋₆alkyl)₂, optionally substituted carbocyclyl, aryl [optionally     substituted with one, two, or three substituents independently     selected from hydroxy, C₁₋₆alkyloxy, halo, haloC₁₋₆alkyl,     haloC₁₋₆alkyloxy, cyano, —NH₂, —NH(C₁₋₆alkyl), and —N(C₁₋₆alkyl)₂],     heteroaryl [optionally substituted with one, two, or three     substituents independently selected from hydroxy, C₁₋₆alkyloxy,     halo, haloC₁₋₆alkyl, haloC₁₋₆alkyloxy, cyano, —NH₂, —NH(C₁₋₆alkyl),     —N(C₁₋₆alkyl)₂], and heterocyclyl optionally substituted with one,     two, or three substituents independently selected from hydroxy,     C₁₋₆alkyloxy, halo, haloC₁₋₆alkyl, haloC₁₋₆alkyloxy, cyano, —NH₂,     —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂]; optionally substituted carbocyclyl     C₁₋₆alkyl is optionally substituted carbocyclyl attached via     C₁₋₆alkyl; -   optionally substituted heterocyclylC₁₋₆alkyl is optionally     substituted heterocyclyl attached via C₁₋₆alkyl; -   optionally substituted arylC₁₋₆alkyl is optionally substituted aryl     attached via C₁₋₆alkyl; and -   optionally substituted heteroarylC₁₋₆alkyl is optionally substituted     heteroaryl attached via C₁₋₆alkyl.

Embodiment 1

In embodiment 1, the compound of Formula (I) is as defined in the Summary.

In a subembodiment of embodiment 1, the compounds of Formula (I) are those wherein when R⁵ is hydrogen, R^(1a) and R^(1b) are independently hydrogen, hydroxy, halo, C₁-C₃alkyl, C₁-C₃alkoxy, C₂-C₃alkenyl, or C₂-C₃alkynyl, R^(2a), R^(2b) and R³ are hydrogen, then Het is not 2,4-dioxo-3,4-dihydropyrimidinyl, 2,4-dioxo-3,4,5,6-tetrahydropyrimidinyl, 6-oxopurinyl, 6-amino-9H-purin-9-yl, or oxopyrimidinyl wherein each ring of the aforementioned ring is optionally substituted with C₁₋₆alkyl.

Embodiment 2

In embodiment 2, the compounds of embodiment 1 and subembodiments contained therein are those wherein R⁵ is phenylC₁₋₆alkyl or naphthylC₁₋₆alkyl, preferably benzyl, wherein phenyl and naphthyl are optionally substituted with one, two, or three substituents independently selected from —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, —C₂₋₆alkenylene-CO₂H, —OC₁₋₆alkylene-CO₂H, —OC₁₋₆alkylene-CO₂C₁₋₆alkyl, —C₁₋₆alkyl substituted with one or two hydroxy, —OC₁₋₆alkyl substituted with one or two hydroxy, —C₁₋₆alkyl substituted with one or two —OC₁₋₆alkyl, —OC₁₋₆alkyl (substituted with one or two —OC₁₋₆alkyl), —CO₂H, —COOC₁₋₆alkyl, hydroxy, halo, nitro, —PO₃H₂, cyano, —NH₂, —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂, —CONR^(e)R^(f) (where R^(e) and R^(f) are independently hydrogen, —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, or —C₁₋₆alkylene-substituted with one or two substituents independently selected from hydroxy, NH₂, —NHC₁₋₆alkyl or —N(C₁₋₆alkyl)₂), —SO₂(C₁₋₆alkyl), —SO₂NR^(g)R^(h) (where R^(g) and R^(h) are independently hydrogen, —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, or —C₁₋₆alkylene-CO₂C₁₋₆alkyl), —SC₁₋₆alkyl, —SOC₁₋₆alkyl, —SO₂NHCOR^(j) (where R^(j) is —C₁₋₆alkyl, —NHC₁₋₆alkyl, or —N(C₁₋₆alkyl)₂), phenyl, —C₁₋₆ alkylenephenyl, phenoxy, —OC₁₋₆alkylenephenyl, 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S [wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl (substituted with one to five fluoro), —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂], —OR, —C₁₋₆alkylene-R^(i), —OC₁₋₆alkylene-R^(i), —SR^(i), —SC₁₋₆alkylene-R^(i), heterocyclyl, —C₁₋₆ alkyleneheterocyclyl, —OC₁₋₆alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m)C₁₋₆alkyleneheterocyclyl, —NR^(m)COC₁₋₆alkyleneNR^(o)R^(p), —NR^(m)COheterocyclyl, —NR^(m)COC₁₋₆alkyleneheterocyclyl, —COheterocyclyl, —CONR^(m)R^(i), —CONR^(m)C₁₋₆alkylene-R^(i), —OCONR^(m)R^(m), —NR^(m)—COR^(y), —NR^(m)—CO—NR^(m)R^(y), —NR^(m)—SO₂—R^(y), —NR^(m)—SO₂—NR^(m)R^(y), and —CONHSO₂R^(z); wherein:

each R^(m) is hydrogen or —C₁₋₆alkyl;

R^(o), R^(p), and R^(y) are independently hydrogen or —C₁₋₆alkyl;

R^(z) is —C₁₋₆alkyl, —NHC₁₋₆alkyl, or —N(C₁₋₆alkyl)₂;

phenyl by itself or as part of —C₁₋₆alkylenephenyl, phenoxy, or —OC₁₋₆alkylenephenyl is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂;

each R^(i) is independently 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S and wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, and —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂;

heterocyclyl by itself or as part of —C₁₋₆alkyleneheterocyclyl, —OC₁₋₆alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m)C₁₋₆alkyleneheterocyclyl, —NR^(m)COC₁₋₆alkyleneheterocyclyl, —COheterocyclyl, or —NR^(m)COheterocyclyl is optionally substituted with one, two, or three substituents independently selected from hydroxy, halo, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, or —C₁₋₆alkyl (substituted with hydroxy or —OC₁₋₆alkyl).

Embodiment 3

In embodiment 3, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is phenylC₁₋₆alkyl or naphthylC₁₋₆alkyl, preferably benzyl, wherein phenyl and naphthyl (also referred to below as R⁵ phenyl and naphthyl rings respectively) are optionally substituted, preferably substituted, with one or two substituents independently selected from —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl substituted with one to five fluoro, —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, —OC₁₋₆alkylene-CO₂H, —OC₁₋₆alkylene-CO₂C₁₋₆alkyl, —C₁₋₆alkyl substituted with one or two hydroxy, —OC₁₋₆alkyl substituted with one or two hydroxy, —C₁₋₆alkyl substituted with one or two substituents independently selected from —OC₁₋₆alkyl, —OC₁₋₆alkyl (substituted with one or two —OC₁₋₆alkyl), —CO₂H, —COOC₁₋₆alkyl, halo, hydroxy, nitro, —PO₃H₂, cyano, —NH₂, —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂, —CONR^(e)R^(f) (where R^(e) and R^(f) are independently hydrogen, —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, or —C₁₋₆alkyl substituted with one or two substituents independently selected from hydroxyl, NH₂, —NHC₁₋₆alkyl or —N(C₁₋₆alkyl)₂), —SO₂(C₁₋₆alkyl), —SO₂NR^(g)R^(h) (where R^(g) and R^(h) are independently hydrogen, —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, or —C₁₋₆alkylene-CO₂C₁₋₆alkyl), —SC₁₋₆alkyl, —SOC₁₋₆alkyl, —SO₂NHCOR^(j) (where R^(j) is —C₁₋₆alkyl, —NHC₁₋₆alkyl, or —N(C₁₋₆alkyl)₂), phenyl, phenoxy, benzyl, —OC₁₋₆alkylenephenyl, 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S [wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂], —OR^(i), —C₁₋₆alkylene-R^(i), —OC₁₋₆alkylene-R^(i), —SR^(i), —SC₁₋₆alkylene-R^(i), heterocyclyl, —C₁₋₆alkyleneheterocyclyl, —OC₁₋₆alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m)C₁₋₆alkyleneheterocyclyl, —NR^(m)COC₁₋₆alkyleneNR^(o)R^(p), —NR^(m)CO-heterocyclyl, —NR^(m)COC₁₋₆alkyleneheterocyclyl, —COheterocyclyl, —CONR^(m)R^(i), —CONR^(m)alkylene-R^(i), and —CONHSO₂R^(z); and

additionally the R⁵ phenyl and naphthyl rings are optionally substituted with a third substituent selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy; wherein:

each R^(m) is hydrogen or —C₁₋₆alkyl;

R^(o) and R^(p) are independently hydrogen or —C₁₋₆alkyl;

R^(z) is —C₁₋₆alkyl, —NHC₁₋₆alkyl, or —N(C₁₋₆alkyl)₂;

phenyl by itself or as part of phenoxy, benzyl, or —OC₁₋₆alkylenephenyl is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂;

each R^(i) is independently 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S and wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, and —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂;

heterocyclyl by itself or as part of —C₁₋₆alkyleneheterocyclyl, —OC₁₋₆alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m)C₁₋₆alkylene-heterocyclyl, —NR^(m)COheterocyclyl, —COheterocyclyl, or —NR^(m)COC₁₋₆alkyleneheterocyclyl is optionally substituted with one, two, or three substituents independently selected from hydroxy, halo, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, or —C₁₋₆alkyl (substituted with hydroxy or —OC₁₋₆alkyl).

Embodiment 4

In embodiment 4, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is phenylC₁₋₆alkyl, preferably benzyl, wherein phenyl is substituted with one substituent at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —C₁₋₆alkyl in —C₁₋₆alkylphenyl or —CH₂— in benzyl respectively, which one substituent is selected from —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, —OC₁₋₆alkylene-CO₂H, —OC₁₋₆alkylene-CO₂C₁₋₆alkyl, —CO₂H, —COOC₁₋₆alkyl, halo, hydroxy, cyano, nitro, —PO₃H₂, —CONR^(e)R^(f) (where R^(e) and R^(f) are independently hydrogen, —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, or —C₁₋₆alkyl substituted with one or two substituents independently selected from hydroxy, NH₂, —NHC₁₋₆alkyl or —N(C₁₋₆alkyl)₂), —SO₂(C₁₋₆alkyl), —SO₂NR^(g)R^(h) (where R^(g) and R^(h) are independently hydrogen, —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, or —C₁₋₆alkylene-CO₂C₁₋₆alkyl), phenyl, phenoxy, benzyl, benzyloxy, 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S [wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxy, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂], —OR^(i), —C₁₋₆alkylene-R^(i), —OC₁₋₆alkylene-R^(i), —SR^(i), —SC₁₋₆alkylene-R^(i), heterocyclyl, —C₁₋₆alkyleneheterocyclyl, —OC₁₋₆alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m)C₁₋₆alkyleneheterocyclyl, —NR^(m)COC₁₋₆alkyleneNR^(o)R^(p), —NR^(m)COheterocyclyl, —NR^(m)COC₁₋₆alkyleneheterocyclyl, —COheterocyclyl, —CONR^(m)R, and —CONR^(m)alkylene-R^(i); and

the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy; wherein

each R^(m) is hydrogen or —C₁₋₆alkyl;

R^(o) and R^(p) are independently hydrogen or —C₁₋₆alkyl;

phenyl by itself or as part of benzyl, phenoxy, or benzyloxy is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆ alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂;

each R^(i) is independently 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S and wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, and —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂;

heterocyclyl by itself or as part of —C₁₋₆alkyleneheterocyclyl, —OC₁₋₆ alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m)C₁₋₆alkyleneheterocyclyl, —NR^(m)COheterocyclyl, —COheterocyclyl, or —NR^(m)COC₁₋₆alkyleneheterocyclyl is optionally substituted with one, two, or three substituents independently selected from hydroxy, halo, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, or —C₁₋₆alkyl (substituted with hydroxy or —OC₁₋₆alkyl).

Embodiment 5

In embodiment 5, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂phenyl or —(CH₂)₂phenyl, preferably —CH₂phenyl (benzyl) wherein phenyl is optionally substituted, preferably substituted, with one substituent at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —C₁₋₆alkyl in —C₁₋₆alkylphenyl and —CH₂— in —CH₂phenyl (benzyl) respectively, which one substituent is selected from —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkyl substituted with hydroxy, —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, —OC₁₋₆alkylene-CO₂H, —OC₁₋₆alkylene-CO₂C₁₋₆alkyl, —CO₂H, nitro, —COOC₁₋₆alkyl, halo, hydroxy, cyano, —CONR^(e)R^(f) (where R^(e) is hydrogen or —C₁₋₆alkyl and R^(f) is —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, or —C₁₋₆alkyl substituted with one or two hydroxyl, —NH₂, —NHC₁₋₆alkyl or —N(C₁₋₆alkyl)₂), —SO₂(C₁₋₆alkyl), —SO₂NR^(g)R^(h) (where R^(g) is hydrogen or —C₁₋₆alkyl and R^(h) is hydrogen, —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, or —C₁₋₆alkylene-CO₂C₁₋₆alkyl), oxadiazolyl, tetrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, oxadiazolyl, thiadiazolyl, furanyl, thienyl, pyrrolyl or isoxazolyl (wherein each of the aforementioned heteroaryl rings is optionally substituted with one or two substituents independently selected from C₁₋₆alkyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂), and

the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 6

In embodiment 6, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is phenylC₁₋₆alkyl, preferably benzyl, wherein phenyl ring in phenylC₁₋₆alkyl and benzyl is substituted at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —C₁₋₆alkyl in —C₁₋₆alkylphenyl and —CH₂— in benzyl respectively, with phenyl, benzyl, benzyloxy, or phenoxy [wherein the phenyl ring either by itself or as part of benzyl, benzyloxy, and phenoxy is optionally substituted with one, two, or three substituents independently selected from hydroxy, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl (substituted with one to five fluoro), —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂]; and

R⁵ phenyl is further optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 7

In embodiment 7, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl, —CH₂-naphthyl, —(CH₂)₂-phenyl, or —(CH₂)₂-naphthyl, preferably —CH₂-phenyl, wherein the phenyl and naphthyl rings are optionally substituted with one, two, or three substituents independently selected from methyl, ethyl, methoxy, ethoxy, difluoromethyl, trifluoromethyl, difluoromethoxy, trifluoromethoxy, —CH₂CO₂H, —CH₂CH₂CO₂H, —OCH₂CO₂H, —OCH₂CH₂CO₂H, —CH₂CO₂CH₃, —CH₂CH₂CO₂CH₃, —CH₂CO₂ethyl, —CH₂CH₂CO₂ethyl, —CH₂OH, —CH₂CH₂OH, —OCH₂OCH₃, —OCH₂CH₂OCH₃, —CO₂H, —COOmethyl, —CO₂ethyl, hydroxy, nitro, PO₃H₂, methylthio, ethylthio, methylsulfoxide, ethylsulfoxide, methylcarbonylaminosulfonyl, ethylcarbonylaminosulfonyl, fluoro, chloro, cyano, —NH₂, —NHCH₃, —NHethyl, —N(methyl)₂, —N(ethyl)₂, —CONR^(e)R^(f) (where R^(e) is hydrogen, methyl, ethyl, or propyl and R^(f) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, —(CH₂)₃—CO₂ethyl, —(CH₂)₂—NH₂, —(CH₂)₃—NH₂, —(CH₂)₂—NHMe, —(CH₂)₃—NHMe, —(CH₂)₂—N(CH₃)₂, or —(CH₂)₃—N(CH₃)₂), —SO₂Me, —SO₂NR^(g)R^(h) (where R^(g) is hydrogen, methyl, ethyl, or propyl and R^(h) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, or —(CH₂)₃—CO₂ethyl), phenyl, phenoxy, —CH₂-phenyl, —CH₂—CH₂-phenyl, —OCH₂-phenyl or —OCH₂—CH₂-phenyl [wherein the phenyl ring, by itself or as part of phenoxy, —CH₂-phenyl, —CH₂—CH₂-phenyl, —OCH₂-phenyl, and —OCH₂—CH₂-phenyl, is optionally substituted with one, two, or three substituents independently selected from hydroxyl, chloro, fluoro, —CO₂H, CN, —CO₂Me, —CO₂ethyl, methyl, ethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, difluoromethyl, trifluoromethyl, —CH₂—CO₂H, —CH₂—CO₂methyl, —CH₂—CO₂ethyl, tetrazolyl, —SO₂CH₃, —SO₂NH₂, —SO₂NHCH₃, —SO₂N(CH₃)₂, —CONH₂, —CONHCH₃, and —CON(CH₃)₂], 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S [wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, chloro, fluoro, CN, —CO₂H, —CO₂Me, —CO₂ethyl, methyl, ethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, difluoromethyl, trifluoromethyl, —CH₂—CO₂H, —CH₂—CO₂methyl, and —CH₂—CO₂ethyl], —OR^(i), —SR^(i), —OCH₂R^(i), —O(CH₂)₂—R^(i), —CH₂R^(i) and —(CH₂)₂—R^(i) [where each R^(i) is independently 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S and wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, chloro, fluoro, CN, —CO₂H, —CO₂Me, —CO₂ethyl, methyl, ethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, difluoromethyl, trifluoromethyl, —CH₂—CO₂H, —CH₂—CO₂methyl, —CH₂—CO₂ethyl, tetrazolyl, —SO₂CH₃, —SO₂NH₂, —SO₂NHCH₃, —SO₂N(CH₃)₂, —CONH₂, —CONHCH₃, and —CON(CH₃)_(2].)

Embodiment 8

In embodiment 8, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl, —CH₂-naphthyl, —(CH₂)₂-phenyl, or —(CH₂)₂-naphthyl, preferably —CH₂-phenyl, wherein phenyl and naphthyl rings are substituted with one or two substituents independently selected from difluoromethyl, trifluoromethyl, difluoromethoxy, trifluoromethoxy, —CH₂CO₂H, —CH₂CH₂CO₂H, —OCH₂CO₂H, —OCH₂CH₂CO₂H, —CH₂CO₂CH₃, —CH₂CH₂CO₂CH₃, —CH₂CO₂ethyl, —CH₂CH₂CO₂ethyl, —CH₂OH, —CH₂CH₂OH, —OCH₂OCH₃, —OCH₂CH₂OCH₃, —CO₂H, —COOmethyl, —CO₂ethyl, hydroxy, nitro, PO₃H₂, methylthio, ethylthio, methylsulfoxide, ethylsulfoxide, methylcarbonylaminosulfonyl, ethylcarbonylaminosulfonyl, fluoro, chloro, cyano, —NH₂, —NHCH₃, —NHethyl, —N(methyl)₂, —N(ethyl)₂, —CONR^(e)R^(f) (where R^(e) is hydrogen, methyl, ethyl, or propyl and R^(f) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, —(CH₂)₃—CO₂ethyl, —(CH₂)₂—NH₂, —(CH₂)₃—NH₂, —(CH₂)₂—NHMe, —(CH₂)₃—NHMe, —(CH₂)₂—N(CH₃)₂, or —(CH₂)₃—N(CH₃)₂), —SO₂Me, —SO₂NR^(g)R^(h) (where R^(g) is hydrogen, methyl, ethyl, or propyl and R^(h) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, or —(CH₂)₃—CO₂ethyl), phenyl, phenoxy, —CH₂-phenyl, —CH₂—CH₂-phenyl, —OCH₂-phenyl or —OCH₂—CH₂-phenyl [wherein the phenyl ring by itself or as part of phenoxy, —CH₂-phenyl, —CH₂—CH₂-phenyl, —OCH₂-phenyl, and —OCH₂—CH₂-phenyl is optionally substituted with one, two, or three substituents independently selected from hydroxyl, chloro, fluoro, —CO₂H, CN, —CO₂Me, —CO₂ethyl, methyl, ethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, difluoromethyl, trifluoromethyl, —CH₂—CO₂H, —CH₂—CO₂methyl, —CH₂—CO₂ethyl, tetrazolyl, —SO₂CH₃, —SO₂NH₂, —SO₂NHCH₃, —SO₂N(CH₃)₂, —CONH₂, —CONHCH₃, and —CON(CH₃)_(2], 5)- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S [wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, chloro, fluoro, CN, —CO₂H, —CO₂Me, —CO₂ethyl, methyl, ethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, difluoromethyl, trifluoromethyl, —CH₂—CO₂H, —CH₂—CO₂methyl, —CH₂—CO₂ethyl, tetrazolyl, —SO₂CH₃, —SO₂NH₂, —SO₂NHCH₃, —SO₂N(CH₃)₂, —CONH₂, —CONHCH₃, and —CON(CH₃)₂], —OR^(i), —SR^(i), —OCH₂R^(i), —O(CH₂)₂—R^(i), —CH₂R^(i) and —(CH₂)₂—R^(i) [where each R^(i) is independently 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S and wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, chloro, fluoro, CN, —CO₂H, —CO₂Me, —CO₂ethyl, methyl, ethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, difluoromethyl, trifluoromethyl, —CH₂—CO₂H, —CH₂—CO₂methyl, and —CH₂—CO₂ethyl]; and

the R⁵ phenyl and naphthyl rings are additionally optionally substituted with a third substituent independently selected from methyl, ethyl, fluoro, chloro, methoxy, ethoxy, hydroxy, —NH₂, and cyano.

Embodiment 9

In embodiment 9, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl, wherein phenyl is substituted with one substituent at a meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl or —(CH₂)₂-phenyl respectively, and is selected from trifluoromethyl, difluoromethoxy, trifluoromethoxy, —CH₂CO₂H, —CH₂CH₂CO₂H, —OCH₂CO₂H, —OCH₂CH₂CO₂H, —CH₂CO₂CH₃, —CH₂CH₂CO₂CH₃, —CH₂CO₂ethyl, —CH₂CH₂CO₂ethyl, —CH₂OH, —CH₂CH₂OH, —OCH₂CH₂OH, —OCH₂OCH₃, —OCH₂CH₂OCH₃, —CO₂H, —COOmethyl, —COOethyl, hydroxy, fluoro, chloro, cyano, —CONR^(e)R^(f) (where R^(e) is hydrogen, methyl, ethyl, or propyl and R^(f) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, —(CH₂)₃—CO₂ethyl, —(CH₂)₂—NH₂, —(CH₂)₃—NH₂, —(CH₂)₂—NHMe, —(CH₂)₃—NHMe, —(CH₂)₂—NHCH₃, —(CH₂)₃—N(CH₃)₂), —SO₂Me, or —SO₂NR^(g)R^(h) (where R^(g) is hydrogen, methyl, ethyl, or propyl and R^(h) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, or —(CH₂)₃—CO₂ethyl), tetrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, oxadiazolyl, thiadiazolyl, furanyl, thienyl, pyrrolyl or isoxazolyl, wherein each of the aforementioned heteroaryl rings is optionally substituted with one or two substituents independently selected from C₁₋₆alkyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, or —OC₁₋₆alkyl (substituted with one to five fluoro); and

the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from methyl, ethyl, fluoro, chloro, methoxy, ethoxy, hydroxy, —NH₂, difluoromethyl, trifluoromethyl, difluoromethoxy, trifluoromethoxy, and cyano.

Embodiment 10

In embodiment 10, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl, wherein the phenyl ring in —CH₂-phenyl and —(CH₂)₂-phenyl is substituted at the meta or para position to the carbon atom of the phenyl ring that is attached to the —CH₂— group of the —CH₂-phenyl and —(CH₂)₂-phenyl ring with haloC₁₋₆alkoxy or haloC₁₋₆alkyl and the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 11

In embodiment 11, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl, wherein phenyl is substituted with a substituent at the meta or para position, preferably para position, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl or —(CH₂)₂-phenyl, which substituent is selected from —CONR^(e)R^(f)(where R^(e) is hydrogen or —C₁₋₆alkyl and R^(f) is —C₁₋₆alkylene-substituted with one or two substituents independently selected from hydroxyl, NH₂, —NHC₁₋₆alkyl or —N(C₁₋₆alkyl)₂), —OR^(i), —C₁₋₆alkylene-R^(i), —OC₁₋₆alkylene-R^(i), —SR^(i), —SC₁₋₆alkylene-R^(i), —CONR^(m)R^(i), —CONR^(m)C₁₋₆alkylene-R^(i) [where each R^(i) is independently 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S and wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), or —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), heterocyclyl, —C₁₋₆alkyleneheterocyclyl, —OC₁₋₆alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m) C₁₋₆alkyleneheterocyclyl, —NR^(m)COheterocyclyl, —COheterocyclyl, —NR^(m)COC₁₋₆alkyleneheterocyclyl [wherein each R^(m) is hydrogen or —C₁₋₆alkyl and the heterocyclyl ring by itself or as part of —C₁₋₆alkyleneheterocyclyl, —OC₁₋₆alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m)C₁₋₆alkyleneheterocyclyl, —COheterocyclyl, —NR^(m)COheterocyclyl, and —NR^(m)COC₁₋₆alkyleneheterocyclyl is optionally substituted with one, two, or three substituents independently selected from hydroxy, halo, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, or —C₁₋₆alkyl (wherein alkyl is substituted with hydroxy or —OC₁₋₆alkyl)], and —NR^(m)COC₁₋₆alkyleneNR^(o)R^(p) (where R^(m), R^(o) and R^(p) are independently hydrogen or C₁₋₆alkyl); and

the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 12

In embodiment 12, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl, wherein phenyl is substituted with —CH═CHCO₂H, —CH₂CO₂H or CH₂CH₂CO₂ at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl or —(CH₂)₂-phenyl; and

the phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 13

In embodiment 13, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl, wherein phenyl is substituted —CH₂CO₂CH₃, —CH₂CH₂CO₂CH₃, —CH₂CO₂ethyl, or —CH₂CH₂CO₂ethyl at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl or —(CH₂)₂-phenyl; and

the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 14

In embodiment 14, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl (benzyl), wherein phenyl is substituted with CO₂H at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl (benzyl) or —(CH₂)₂-phenyl and the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 15

In embodiment 15, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl (benzyl) or —(CH₂)₂-phenyl, preferably —CH₂-phenyl (benzyl), wherein phenyl is substituted with —COOmethyl or —COOethyl at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl (benzyl) or —(CH₂)₂-phenyl and the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 16

In embodiment 16, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl, wherein phenyl is substituted with fluoro, chloro, or cyano at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl or —(CH₂)₂-phenyl and the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy

Embodiment 17

In embodiment 17, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl, wherein phenyl is substituted with —CONR^(e)R^(f) (where R^(e) is hydrogen, methyl, ethyl, or propyl and R^(f) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, —(CH₂)₃—CO₂ethyl, —(CH₂)₂—NH₂, —(CH₂)₃—NH₂, —(CH₂)₂—NHMe, —(CH₂)₃—NHMe, —(CH₂)₂—NHCH₃, —(CH₂)₃—N(CH₃)₂) or —CONHSO₂R^(z) where R^(z) is —C₁₋₆alkyl, —NHC₁₋₆alkyl, or —N(C₁₋₆alkyl)₂ at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl or —(CH₂)₂-phenyl; and

the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 18

In embodiment 18, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl, wherein phenyl is substituted —SO₂NR^(g)R^(h) (where R^(g) is hydrogen, methyl, ethyl, or propyl and R^(h) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, or —(CH₂)₃—CO₂ethyl) or —SO₂NHCOR^(j) (where R^(j) is —C₁₋₆alkyl, —NHC₁₋₆alkyl, or —N(C₁₋₆alkyl)₂) at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl or —(CH₂)₂-phenyl; and

the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 19

In embodiment 19, the compounds of embodiment 1 or 2 and subembodiments contained therein are those wherein R⁵ is —CH₂-phenyl or —(CH₂)₂-phenyl, preferably —CH₂— phenyl, wherein phenyl is substituted with tetrazol-5-yl at the meta or para position, preferably para, to the carbon atom of the phenyl ring that is attached to —CH₂— or —(CH₂)₂— in —CH₂-phenyl or —(CH₂)₂-phenyl; and

the R⁵ phenyl ring is additionally optionally substituted with one or two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 20

In embodiment 20, the compounds of embodiment 1 and subembodiments contained therein are those wherein R⁵ is 5-10 membered heteroarylC₁₋₆alkyl (also referred to below as R⁵ heteroaryl), preferably —CH₂— or —(CH₂)₂-5-10 membered heteroaryl, having one to three heteroatoms independently selected from N, O, or S and is optionally substituted, preferably substituted, with one or two substituents, more preferably one substituent, independently selected from —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆ alkyl substituted with one to five fluoro, —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, —C₂₋₆alkenylene-CO₂H, —C₁₋₆alkyl substituted with one or two substituents independently selected from hydroxy, —OC₁₋₆alkyl substituted with hydroxy, —C₁₋₆alkyl substituted with one or two —OC₁₋₆alkyl, —OC₁₋₆alkyl (substituted with one or two —OC₁₋₆alkyl), —CO₂H, —COOC₁₋₆ alkyl, hydroxy, halo, nitro, —PO₃H₂, cyano, —NH₂, —NH(C₁₋₆alkyl), —N(C₁₋₆alkyl)₂, —CONR^(e)R^(f) (where R^(e) and R^(f) are independently hydrogen, —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, or —C₁₋₆alkyl substituted with one or two substituents independently selected from hydroxyl, —NHC₁₋₆alkyl or —N(C₁₋₆alkyl)₂), —SO₂(C₁₋₆alkyl), —SO₂NR^(g)R^(h) (where R^(g) and R^(h) are independently hydrogen, —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, or —C₁₋₆alkylene-CO₂C₁₋₆alkyl), —SC₁₋₆alkyl, —SOC₁₋₆alkyl, —SO₂NHCOR^(j) (where R^(j) is —C₁₋₆alkyl, —NH₂, —NHC₁₋₆alkyl, or —N(C₁₋₆alkyl)₂), phenyl, —C₁₋₆alkylenephenyl, phenoxy, —OC₁₋₆alkylenephenyl, 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S [wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl (substituted with one to five fluoro), —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂], —OR^(i), —C₁₋₆alkylene-R^(i), —OC₁₋₆alkylene-R^(i), —SR^(i), —SC₁₋₆alkylene-R^(i), heterocyclyl, —C₁₋₆alkyleneheterocyclyl, —OC₁₋₆alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m)C₁₋₆alkyleneheterocyclyl, —NR^(m)COC₁₋₆alkyleneNR^(o)R^(p), —NR^(m)COheterocyclyl, —NR^(m)COC₁₋₆alkyleneheterocyclyl, —COheterocyclyl, —CONR^(m)R^(i), —CONR^(m)alkylene-R^(i), —OCONR^(m)R^(m), —NR^(m)—COR^(y), —NR^(m)—CO—NR^(m)R^(y), —NR^(m)—SO₂—R^(y), —NR^(m)—SO₂—NR^(m)R^(y), and —CONHSO₂R^(z); and

the R⁵ heteroaryl ring is additionally optionally substituted with an additional substituent selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy; wherein:

each R^(m) is hydrogen or —C₁₋₆alkyl;

R^(o), R^(p), and R^(y) are independently hydrogen or —C₁₋₆alkyl;

R^(z) is —C₁₋₆alkyl, —NHC₁₋₆alkyl, or —N(C₁₋₆alkyl)₂;

phenyl by itself or as part of —C₁₋₆alkylenephenyl, phenoxy, or —OC₁₋₆alkylenephenyl is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂;

each R^(i) is independently 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S and wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, and —C₁₋₆alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂;

heterocyclyl by itself or as part of —C₁₋₆alkyleneheterocyclyl, —OC₁₋₆ alkyleneheterocyclyl, —SC₁₋₆alkyleneheterocyclyl, —CONR^(m)C₁₋₆alkyleneheterocyclyl, —NR^(m)COheterocyclyl, —COheterocyclyl, and —NR^(m)COC₁₋₆alkyleneheterocyclyl is optionally substituted with one, two, or three substituents independently selected from hydroxy, halo, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, or —C₁₋₆alkyl (substituted with hydroxy or —OC₁₋₆alkyl).

Embodiment 21

In embodiment 21, the compounds of embodiment 1 or 20 and subembodiments contained therein are those wherein R⁵ is —CH₂— or —(CH₂)₂-(5-9 membered heteroaryl ring) which heteroaryl ring is optionally substituted with one or two substituents, preferably one substituent, independently selected from difluoromethyl, trifluoromethyl, difluoromethoxy, trifluoromethoxy, —CH₂CO₂H, —CH₂CH₂CO₂H, —CH₂CO₂CH₃, —CH₂CH₂CO₂CH₃, —CH₂CO₂ethyl, —CH₂CH₂CO₂ethyl, —CH₂OH, —CH₂CH₂OH, —OCH₂CH₂OH, —OCH₂OCH₃, —OCH₂CH₂OCH₃, —CO₂H, —COOmethyl, —CO₂ethyl, hydroxy, nitro, PO₃H₂, methylthio, ethylthio, methylsulfoxide, ethylsulfoxide, methylcarbonylaminosulfonyl, ethylcarbonylaminosulfonyl, fluoro, chloro, cyano, —NH₂, —NHCH₃, —NHethyl, —N(methyl)₂, —N(ethyl)₂, —CONR^(e)R^(f) (where R^(e) is hydrogen, methyl, ethyl, or propyl and R^(f) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, —(CH₂)₃—CO₂ethyl, —(CH₂)₂—NH₂, —(CH₂)₃—NH₂, —(CH₂)₂—NHMe, —(CH₂)₃—NHMe, —(CH₂)₂—N(CH₃)₂, or —(CH₂)₃—N(CH₃)₂), —SO₂Me, —SO₂NR^(g)R^(h) (where R^(g) is hydrogen, methyl, ethyl, or propyl and R^(h) is hydrogen, methyl, ethyl, propyl, —(CH₂)₂—CO₂H, —(CH₂)₃—CO₂H, —(CH₂)₂—CO₂Me, —(CH₂)₃—CO₂Me, —(CH₂)₂—CO₂ethyl, or —(CH₂)₃—CO₂ethyl), phenyl, phenoxy, —CH₂-phenyl, —CH₂—CH₂-phenyl, —OCH₂-phenyl or —OCH₂—CH₂-phenyl [wherein the phenyl ring by itself or as part of phenoxy, —CH₂-phenyl, —CH₂—CH₂-phenyl, —OCH₂-phenyl, and —OCH₂—CH₂-phenyl is optionally substituted with one, two, or three substituents independently selected from hydroxyl, chloro, fluoro, —CO₂H, CN, —CO₂Me, —CO₂ethyl, methyl, ethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, difluoromethyl, trifluoromethyl, —CH₂—CO₂H, —CH₂—CO₂methyl, —CH₂—CO₂ethyl, tetrazolyl, —SO₂CH₃, —SO₂NH₂, —SO₂NHCH₃, —SO₂N(CH₃)₂, —CONH₂, —CONHCH₃, and —CON(CH₃)₂], 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from 0, N, and S [wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, chloro, fluoro, CN, —CO₂H, —CO₂Me, —CO₂ethyl, methyl, ethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, difluoromethyl, trifluoromethyl, —CH₂—CO₂H, —CH₂—CO₂methyl, —CH₂—CO₂ethyl, tetrazolyl, —SO₂CH₃, —SO₂NH₂, —SO₂NHCH₃, —SO₂N(CH₃)₂, —CONH₂, —CONHCH₃, and —CON(CH₃)₂], —OR^(i), —SR^(i), —OCH₂R^(i), —O(CH₂)₂—R^(i), —CH₂R and —(CH₂)₂R^(i) [where each R^(i), is independently 5- or 6-membered monocyclic heteroaryl containing one to four heteroatoms independently selected from O, N, and S and wherein the heteroaryl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, chloro, fluoro, CN, —CO₂H, —CO₂Me, —CO₂ethyl, methyl, ethyl, methoxy, ethoxy, difluoromethoxy, trifluoromethoxy, difluoromethyl, trifluoromethyl, —CH₂—CO₂H, —CH₂—CO₂methyl, and —CH₂—CO₂ethyl]; and

the R⁵ heteroaryl ring is optionally additionally substituted with a substituent selected from methyl, ethyl, fluoro, chloro, methoxy, ethoxy, hydroxy, —NH₂, and cyano.

Embodiment 22

In embodiment 22, the compounds of embodiment 1 or 20 and subembodiments contained therein are those wherein R⁵ is —(CH₂)₁₋₂-(5-10 membered heteroaryl ring) having one to three heteroatoms independently selected from N, O, or S, preferably —CH₂-(5-9 membered heteroaryl ring) having one to 3 heteroatoms independently selected from N, O, or S, wherein the heteroaryl ring is substituted with one or two substituents independently selected from —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆alkylene-CO₂C₁₋₆alkyl, —CO₂H, nitro, —COOC₁₋₆alkyl, halo, hydroxy, cyano, —CONR^(e)R^(f) (where R^(e) is hydrogen or —C₁₋₆alkyl and R^(f) is —C₁₋₆alkyl, —C₁₋₆alkylene-CO₂H, or —C₁₋₆alkylene-CO₂C₁₋₆alkyl), —SO₂(C₁₋₆ alkyl), —SO₂NR^(g)R^(h) (where R^(g) is hydrogen or —C₁₋₆alkyl and R^(h) is hydrogen, —C₁₋₆alkyl, —C₁₋₆ alkylene-CO₂H, or —C₁₋₆alkylene-CO₂C₁₋₆alkyl), tetrazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrazolyl, imidazolyl, oxazolyl, thiazolyl, oxadiazolyl, thiadiazolyl, furanyl, thienyl, pyrrolyl or isoxazolyl, (wherein each of the aforementioned heteroaryl rings is optionally substituted with one or two substituents independently selected from C₁₋₆alkyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl substituted with one to five fluoro, —OC₁₋₆alkyl (substituted with one to five fluoro, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂); and

the R⁵ heteroaryl ring is additionally optionally substituted with one or two substituents, independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 23

In embodiment 23, the compounds of embodiment 1 or 20 and subembodiments contained therein are those wherein R⁵ is 5-10 membered heteroarylC₁₋₆alkyl having one to 3 heteroatoms independently selected from N, O, or S wherein the heteroaryl ring is substituted with phenyl [wherein the phenyl ring is optionally substituted with one, two, or three substituents independently selected from hydroxyl, halo, CN, —CO₂H, —COOC₁₋₆alkyl, —C₁₋₆alkyl, —OC₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —C₁₋₆alkylene-CO₂H, —C₁₋₆ alkylene-CO₂C₁₋₆alkyl, tetrazolyl, —SO₂C₁₋₆alkyl, —SO₂NH₂, —SO₂NHC₁₋₆alkyl, —SO₂N(C₁₋₆alkyl)₂, —CONH₂, —CONHC₁₋₆alkyl, and —CON(C₁₋₆alkyl)₂]; and

the R⁵ heteroaryl ring is further optionally substituted with two substituents independently selected from —C₁₋₆alkyl, —C₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), —O—C₁₋₆alkyl, —OC₁₋₆alkyl (wherein alkyl is substituted with one to five fluoro), halo, CN, —NH₂, and hydroxy.

Embodiment 24

In embodiment 24, the compounds of embodiment 1, 20, 21, 22, or 23 and subembodiments contained therein are those wherein the heteroaryl ring in the —CH₂-(5-10 membered heteroaryl ring) of R⁵ is selected from thienyl, furanyl, pyrazolyl, imidazolyl, triazolyl, pyridinyl, pyrazinyl, pyridazinyl, pyrimidinyl, oxazolyl, oxadiazolyl, isoxazolyl, thiazolyl, thiadiazolyl, benzimidazolyl, benzofuranyl, benzoxazolyl, indolyl, and benzothienyl. In one subembodiment of embodiment 24, the heteroaryl ring is a 9- or 10-membered heteroaryl ring selected from benzimidazol-5-yl, benzoxazol-5-yl, indol-5-yl, benzimidazol-2-yl, benzoxazol-2-yl, benzofuran-5-yl, indol-2-yl, quinolinyl, or isoquinolinyl. In one subembodiment of embodiment 24, the heteroaryl ring is 5-membered heteroaryl ring that is preferably substituted at the 3-position to the ring atom of the 5-membered ring that is attached to —CH₂—, and the 6-membered heteroaryl ring is substituted at a meta or para position to the ring atom that is attached to the —CH₂— and furthermore, the above rings are optionally substituted, preferably substituted, with one or more substituent(s) as indicated in embodiments 20, 21, 22, or 23 above.

Embodiment 25

In embodiment 25, the compounds of any one of embodiments 1-24 and subembodiments contained therein are those wherein R³ is hydrogen, X is O, and Y is O.

Embodiment 26

In embodiment 26, the compounds of any one of embodiments 1-24 and subembodiments contained therein are those are wherein R³ is hydrogen, C₁₋₆alkyl, or benzyl, X is CH₂, and Y is O. In one group of compounds within embodiment 26, R³ is hydrogen, X is CH₂, and Y is O.

Embodiment 27

In embodiment 27, the compounds of any one of embodiments 1-24 and subembodiments contained therein are those wherein R³ is hydrogen, X is N. Within embodiment 27, in one group of compounds Y is O. Within embodiment 27, in another group of compounds Y is S.

Embodiment 28

In embodiment 28, the compounds of any one of embodiments 1-24 and subembodiments contained therein are those wherein R³ is hydrogen, X is O, and Y is S.

Embodiment 29

In embodiment 29, the compounds of any one of embodiments 1-24 and subembodiments contained therein and groups contained therein are those wherein R³ is hydrogen, X is CH₂, and Y is S.

Embodiment 30

In embodiment 30, the compounds of any one of embodiments 1-29 and subembodiments contained therein and groups contained therein are those wherein:

R^(1a) is hydroxy, halo, C₁₋₆alkyl, cyano, azido, NH₂, C₁₋₆alkylcarbonyloxy, ethynyl, or vinyl; preferably hydroxy, fluoro, chloro, cyano, azido, NH₂, or methyl;

R^(1b) is hydrogen;

R^(2a) is hydrogen, hydroxy, halo, C₁₋₆alkyl, NH₂, cyano, azido, ethynyl, or vinyl; and

R^(2b) is hydrogen.

Embodiment 31

In embodiment 31, the compounds of embodiment 30 are those wherein:

R^(1a) is hydroxy, fluoro, or methyl;

R^(1b) is hydrogen;

R^(2a) is hydroxy, fluoro, or methyl; and

R^(2b) is hydrogen.

Embodiment 32

In embodiment 32, the compounds of embodiment 30 are those wherein:

R^(1a) is fluoro;

R^(1b) is hydrogen;

R^(2a) is hydroxy, hydrogen, methyl or NH₂; and

R^(2b) is hydrogen.

Within embodiment 32, in one group of compounds R^(2a) is hydrogen.

Within embodiment 32, in another group of compounds R^(2a) is hydroxy.

Within embodiment 32, in yet another group of compounds R^(2a) is methyl.

Within embodiment 32, in yet another group of compounds R^(2a) is NH₂.

Embodiment 33

In embodiment 33, the compounds of embodiment 30, 31 and 32 and groups contained therein are those wherein

Embodiment 34

In embodiment 34, the compounds of embodiment 30, 31 and 32 and groups contained therein are those wherein:

Embodiment 35

In embodiment 35, the compounds of embodiment 30, 31 and 32 and groups contained therein are those wherein

Embodiment 36

In embodiment 36, the compounds of embodiment 30, 31 and 32 and groups contained therein are those wherein:

Embodiment 37

In embodiment 37, the compounds of embodiment 30, 31 and 32 and groups contained therein are those wherein:

Embodiment 38

In embodiment 38, the compounds of embodiment 30, 31 and 32 and groups contained therein are those wherein:

Embodiment 39

In embodiment 39, the compounds of any one of embodiments 1-29 are those wherein:

R^(1a) is halo, C₁₋₆alkyl, cyano, azido, NH₂, C₁₋₆alkylcarbonyloxy, ethynyl, or vinyl;

R^(1b) is hydroxy or fluoro;

R^(2a) is hydrogen, halo, hydroxy, C₁₋₆alkyl, C₁₋₆alkyloxy, NH₂, ethynyl, or vinyl; and

R^(2b) is hydrogen.

Embodiment 40

In embodiment 40, the compounds of embodiment 39 are those wherein:

R^(1a) is fluoro or methyl;

R^(1b) is fluoro or hydroxy;

R^(2a) is hydrogen, hydroxy, NH₂, methyl, methoxy, ethoxy, or propoxy; and

R^(2b) is hydrogen.

Embodiment 41

In embodiment 41, the compounds of embodiments any one of embodiments 1-29 are those wherein:

R^(1a) is hydroxy, halo, C₁₋₆alkyl, or cyano;

R^(1b) is hydrogen;

R^(2a) is hydroxy, halo, C₁₋₆alkyl, cyano, ethynyl, or vinyl; and

R^(2b) is halo, hydroxy, or C₁₋₆alkyl.

Embodiment 42

In embodiment 42, the compound of embodiment 42 are those wherein:

R^(1a) is hydroxy, fluoro, or methyl;

R^(1b) is hydrogen;

R^(2a) is methyl, fluoro, cyano, or hydroxy; and

R^(2b) is fluoro, hydroxyl, or methyl.

Embodiment 43

In embodiment 43, the compound of embodiment 42 are those wherein:

R^(1a) is fluoro or methyl;

R^(1b) is hydrogen;

R^(2a) is methyl; and

R^(2b) is fluoro, hydroxy, or methyl.

Embodiment 44

In embodiment 44, the compound of any one of embodiments 1-29 and subembodiments and groups of compounds contained therein are those wherein:

R^(1a) is halo, C₁₋₆alkyl, cyano, azido, NH₂, C₁₋₆alkylcarbonyloxy, ethynyl, or vinyl;

R^(1b) is hydroxy or fluoro;

R^(2a) is hydroxy, halo, C₁₋₆alkyl, or cyano; and

R^(2b) is halo, hydroxy, or C₁₋₆alkyl.

Embodiment 45

In embodiment 45, the compounds of any one of embodiments 1 to 45 and subembodiments and groups of compounds contained therein are those wherein:

R⁴ is selected from —C(O)OR⁹, —C(O)NR¹¹R¹², —S(O)₂R¹⁰, —P(O)(OR¹¹)(OR¹²), and —P(O)(OR¹¹)(NR¹³R¹⁵); and

R⁶ is selected from —C(O)OR⁹ and —P(O)(OR¹¹)(OR¹²); wherein: R⁹, R¹⁰, R¹¹, R¹², R¹³ and R¹⁵ are independently hydrogen or C₁₋₆ alkyl.

Within the groups of compounds in embodiment 46, in one group of compounds, R⁴ and R⁶ are —C(O)OR⁹ where R⁹ is as defined above, preferably R⁹ is hydrogen or C₁₋₆ alkyl.

Within the groups of compounds in embodiment 46, in another group of compounds, R⁴ and R⁶ are —C(O)OH.

Within the groups of compounds in embodiment 46, in yet another group of compounds, R⁴ is selected from —C(O)NR¹¹R¹² or —S(O)₂R¹¹; and

R⁶ is selected from —C(O)OR⁹ and —P(O)(OR¹¹)(OR¹²); wherein: R⁹, R¹¹, and R¹² are independently hydrogen or C₁₋₆ alkyl. In one subembodiment, the compounds are those wherein R⁹, R¹¹, and R¹² are independently hydrogen, methyl, or ethyl.

Within the groups of compounds in embodiment 46, in yet another group of compounds, R⁴ is P(O)(OR¹¹)(OR¹²); and

R⁶ is —P(O)(OR¹¹)(OR¹²); wherein:

R¹¹ and R¹² are independently hydrogen or C₁₋₆ alkyl, preferably R¹¹ and R¹² are independently hydrogen, methyl, or ethyl.

Embodiment 46

In embodiment 46, the compounds of embodiments 1 to 46 and subembodiments and groups of compounds contained therein are those wherein: Het is a group selected from a group of formula (i) through (xiv) below:

wherein:

R^(u) is hydrogen, halo, cyano, —NH₂, —NHR²⁰, —NHCOR²⁰, —NR²⁰R²¹, —R²⁰, —SR²⁰, —OH, and —OR²⁰;

R^(w) is hydrogen, halo, —NHR²², —NR²²R²³, —R²², —OH, and —OR²²;

R^(v) and R^(x) are independently hydrogen, halo, haloC₁₋₆alkyl, —NH₂, —NHR²⁴, —NR²⁴R²⁵, —R²⁴, —SR²⁴, cyano, —OH, —OR²⁴, —SO₂R²⁴, —C₁₋₆alkyleneNH₂, —C₁₋₆alkyleneNHR²⁴, —C₁₋₆alkyleneNR²⁴R²⁵, —R²⁴, —C₁₋₆alkyleneSR²⁴, —C₁₋₆alkyleneOH, —C₁₋₆alkyleneOR²⁴, —C₁₋₆alkyleneSO₂R²⁴,

R^(s) and R^(t) are independently hydrogen, halo, or C₁₋₆alkyl; and wherein:

R²⁰, R²¹, R²², R²³, R²⁴ and R²⁵ are independently optionally substituted C₁₋₆alkyl, —C₂-C₆alkenyl, —C₂-C₆alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylC₁₋₆alkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylC₁₋₆alkyl, optionally substituted aryl, optionally substituted arylC₁₋₆alkyl, optionally substituted heteroaryl, or optionally substituted heteroarylC₁₋₆alkyl; or R²⁰ and R²¹, R²² and R²³, and R²⁴ and R²⁵ together with the nitrogen to which they are attached form an optionally substituted nitrogen-containing heterocyclyl.

Embodiment 47

In embodiment 47, the compounds of embodiment 47 are those wherein Het is a group of formula (i).

Embodiment 48

In embodiment 48, the compounds of embodiment 47 are those wherein Het is a group of formula (ii).

Embodiment 49

In embodiment 49, the compounds of embodiment 47 are those wherein Het is a group of formula (iii).

Embodiment 50

In embodiment 50, the compounds of embodiment 47 are those wherein Het is a group of formula (iv).

Embodiment 51

In embodiment 51, the compounds of embodiment 47 are those wherein Het is a group of formula (vi).

Embodiment 52

In embodiment 52, the compounds of embodiment 47 are those wherein Het is a group of formula (vii).

Embodiment 53

In embodiment 53, the compounds of embodiment 47 are those wherein Het is a group of formula (viii).

Embodiment 54

In embodiment 54, the compounds of any one of embodiments 47-54 and subembodiments and groups of compounds contained therein are those wherein R^(s) is hydrogen.

Embodiment 55

In embodiment 55, the compounds of any one of embodiments 47-55 and subembodiments and groups of compounds contained therein are those wherein R^(w) and R^(d)are hydrogen.

Embodiment 56

In embodiment 56, the compounds of any one of embodiments 47-56 and embodiments and groups of compounds contained therein are those wherein: R^(u) is hydrogen, halo, cyano, —NH₂, —NHR²⁰, —NHCOR²⁰, —NR²⁰R²¹, —R²⁰, —SR²⁰, or —OR²⁰ wherein R²⁰ and R²¹ are independently optionally substituted C₁₋₆alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylC₁₋₆alkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylC₁₋₆alkyl, optionally substituted aryl, optionally substituted arylC₁₋₆alkyl, optionally substituted heteroaryl, or optionally substituted heteroarylC₁₋₆alkyl; or R²⁰ and R²¹ together with the nitrogen to which they are attached form an optionally substituted nitrogen-containing heterocyclyl;

R^(v) is hydrogen, halo, haloC₁₋₆alkyl, cyano, —R²⁴, —SR²⁴, —OR²⁴, or —SO₂R²⁴; wherein:

R²⁴ is optionally substituted C₁₋₆alkyl, —C₂-C₆alkenyl, —C₂-C₆alkynyl, optionally substituted cycloalkyl, optionally substituted cycloalkylC₁₋₆alkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylC₁₋₆alkyl, optionally substituted aryl, optionally substituted arylC₁₋₆alkyl, optionally substituted heteroaryl, or optionally substituted heteroarylC₁₋₆alkyl.

Embodiment 57

In embodiment 57, the compounds of any one of embodiments 47-56 and subembodiments and groups of compounds contained therein are those wherein: R^(u) is —NH₂, —NHR²⁰, or —NR²⁰R²¹ wherein R²⁰ and R²¹ are independently optionally substituted C₁₋₆alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylC₁₋₆alkyl, optionally substituted heterocyclyl, optionally substituted heterocyclyl₁₋₆alkyl, optionally substituted aryl, optionally substituted arylC₁₋₆alkyl, optionally substituted heteroaryl, or optionally substituted heteroarylC₁₋₆alkyl; or R²⁰ and R²¹ together with the nitrogen to which they are attached form an optionally substituted nitrogen-containing heterocyclyl; preferably —NH₂, and

R^(v) is halo, preferably chloro.

Representative compounds of Formula (I) include those in Table 1A:

TABLE 1A Cpd. # Compound Name 6

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2- benzylmalonic acid 7

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-(4- carboxybenzyl)-malonic acid 8

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-((6- chloropyridin-3- yl)methyl)malonic acid 10

2-(((2R,3R,4R,5R)-5-(6-amino- 9H-purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-benzylmalonic acid 12

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4-(2- carboxyethyl)benzyl)malonic acid 13

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-2- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- phenoxybenzyl)malonic acid 15

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- (trifluoromethoxy)benzyl)malon- ic acid 16

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- (trifluoromethyl)benzyl)malonic acid 17

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(naphthalen-2- ylmethyl)malonic acid 20

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (trifluoromethyl)benzyl)malonic acid 21

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (methylsulfonyl)benzyl)malonic acid 22

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- (dimethylcarbamoyl)benzyl)ma- lonic acid 23

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(thiophen-3- ylmethyl)malonic acid 25

diethyl 2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 4-fluoro-3- (isobutyryloxy)tetrahydrofuran- 2-yl)methoxy)-2-(4- (trifluoromethoxy)benzyl)malonate 27

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- aminobenzyl)malonic acid 28

2-benzyl-2-(((2R,3R,4S,5R)-5- (6-(benzylamino)-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)malonic acid 32

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-((6- (trifluoromethyl)pyridin-3- yl)methyl)malonic acid 34

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- cyanobenzyl)malonic acid 35

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- cyanobenzyl)malonic acid 36

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (trifluoromethoxy)benzyl)malonic acid 37

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-3-ethoxy-3-oxo-2- (4- (trifluoromethoxy)benzyl)propa- noic acid 38

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-3-ethoxy-3-oxo-2- (4- (trifluoromethoxy)benzyl)propa- noic acid 39

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (hydroxymethyl)benzyl)malonic acid 40

2-(((2R,3R,4S,5R)-5-(6-amino- 2-azido-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- cyanobenzyl)malonic acid 41

2-(((2R,3R,4S,5R)-5-(2-azido- 6-((tert-butoxycarbonyl)amino)- 9H-purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- cyanobenzyl)malonic acid 42

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-(3- carboxybenzyl)-malonic acid 43

2-(3-(1H-tetrazol-5-yl)benzyl)- 2-(((2R,3R,4S,5R)-5-(6-amino- 2-azido-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)malonic acid 44

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- fluorobenzyl)malonic acid 45

diethyl 2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- fluorobenzyl)malonate 46

diethyl 2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- fluroobenzyl)malonate 47

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- fluorobenzyl)malonic acid 48

diethyl 2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (trifluoromethoxy)benzyl)malonate 49

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- methoxybenzyl)malonic acid 50

2-((1H-tetrazol-5-yl)methyl)-2- (((2R,3R,4S,5R)-5-(6-amino-2- chloro-9H-purin-9-yl)-4-fluoro- 3-hydroxytetrahydrofuran-2- yl)methoxy)malonic acid 51

2-(((2S,3S,4R,5R)-3-amino-5- (6-amino-2-chloro-9H-purin-9- yl)-4-hydroxytetrahydrofuran-2- yl)methoxy)-2-benzylmalonic acid 53

2-(3-(1H-tetrazol-5-yl)benzyl)- 2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)malonic acid 54

2-(4-(1H-tetrazol-5-yl)benzyl)- 2-(((2R,3R,4S,5R)-5-(6-amio- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)malonic acid 55

2-(((2R,3S,4R,5R)-4-amino-5- (6-amino-2-chloro-9H-purin-9- yl)-3-hydroxytetrahydrofuran-2- yl)methoxy)-2-benzylmalonic acid 56

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3- hydroxybenzyl)malonic acid 57

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4-carboxy-2- fluorobenzyl)malonic acid 58

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-(4- carboxy-3-fluorobenzyl)malonic acid 59

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-((5- (trifluoromethyl)-furan-2- yl)methyl)malonic acid 60

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-(3- fluoro-4- (trifluoromethyl)benzyl)malonic acid 61

2-(((2R,3R,4S,5R)-5-(6-amnio- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-((3- phenylisoxazol-5- yl)methyl)malonic acid 62

dimethyl 2-(((2R,3R,4S,5R)-5- (6-amino-2-chloro-9H-purin-9- yl)-4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (trifluoromethyl)benzyl)malonate 63

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-((1- benzyl-1H-pyrazol-4- yl)methyl)malonic acid 64

2-((1H-pyrazol-4-yl)methyl)-2- (((2R,3R,4S,5R)-5-(6-amino-2- chloro-9H-purin-9-yl)-4-fluoro- 3-hydroxytetrahydrofuran-2- yl)methoxy)malonic acid 65

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (benzyloxy)benzyl)malonic acid 66

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- hydroxybenzyl)malonic acid 67

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-((2- carboxythiazol-5- yl)methyl)malonic acid 68

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-((2- carboxythiazol-4- yl)methyl)malonic acid 69

2-([1,1′-biphenyl]-4-ylmethyl)- 2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-3-ethoxy- 3-oxopropanoic acid 70

diethyl 2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4-(2-methoxy- 2-oxoethyl)benzyl)malonate 71

diethyl 2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 4-fluoro-3- (isobutyryloxy)tetrahydrofuran- 2-yl)methoxy)-2-((5- (methoxycarbonyl)thiophen-3- yl)methyl)malonate 72

diethyl 2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-((5- (methoxycarbonyl)-1-methyl- 1H-pyrazol-3- yl)methyl)malonate 73

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-((5-carboxy-1- methyl-1H-pyrazol-3- yl)methyl)malonic acid 74

2-benzyl-2-(((2R,3R,4S,5R)-5- (2-chloro-6-hydroxy-9H-purin- 9-yl)-4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)malonic acid 75

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4-cyano-3- fluorobenzyl)malonic acid 76

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-((5- carboxythiophen-3- yl)methyl)malonic acid 77

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-((5- carboxythiopen-2- yl)methyl)malonic acid 78

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (carboxymethyl)benzyl)malonic acid 79

2-([1,1′-biphenyl]-4-ylmethyl)- 2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluroo-3-hydroxytetrahydro- furan-2-yl)methoxy)malonic acid 80

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4-chloro-2- fluorobenzyl)malonic acid 81

2-([1,1′-biphenyl]-4-ylmethyl)- 2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-3-methoxy-3- oxopropanoic acid 82

2-([1,1′-biphenyl]-4-ylmethyl)- 2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-3-methoxy-3- oxopropanoic acid 83

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-((3- carboxyisoxazol-5- yl)methyl)malonic acid 84

2-([1,1′-biphenyl]-4-ylmethyl)- 2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-3-ethoxy- 3-oxopropanoic acid 85

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(3-chloro-4- methoxybenzyl)malonic acid 86

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- sulfamoylbenzyl)malonic acid 87

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-(4-((2- carboxy- ethyl)carbamoyl)benzyl)malonic acid 88

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3-hydroxytetrahydro- furan-2-yl)methoxy)-2-((2- carboxybenzofuran-5- yl)methyl)malonic acid 89

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4-(2,2,2- trifluoroethoxy)benzyl)malonic acid 90

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4-((E)-2- carboxyvinyl)benzyl)malonic acid 91

2-(((2R,3R,4S,5R)-5-(6-amio- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (methoxycarbonyl)benzyl)malonic acid 92

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (carboxymethoxy)benzyl)malonic acid 93

2-benzyl-2-(((2R,3R,4S,5R)-5- (2-chloro-6-oxo-1H-purin- 9(6H)-yl)-4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)malonic acid 94

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- y)methoxy)-2-(4-(5-methyl- 1,3,4-oxadiazol-2- yl)benzyl)malonic acid 95

of 2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-((2′-cyano-[1,1′- biphenyl]-4-yl)-methyl)malonic acid 96

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-((5- chlorobenzo[b]thiophen-3- yl)methyl)-malonic acid 97

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3 hydroxytetrahydrofuran-2- yl)methoxy)-2-(benzo[d]thiazol- 2-ylmethyl)malonic acid 98

2-(((2R,3R,4S,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-4- fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-(4- (methylcarbamoyl)benzyl)malonic acid 99

Diethyl 2-(((2R,3S,4R,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 3,4-dihydroxytetrahydrofuran- 2-yl)methoxy)-2-benzyl malonate 100

2-(((2R,3S,4R,5R)-5-(6-amino- 2-chloro-9H-purin-9-yl)-3,4- dihydroxytetrahydrofuran-2- yl)methoxy)-2-(4- carboxybenzyl)malonic acid 101

S)-2-(((2R,3S,4R,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 3,4-dihydroxytetrahydrofuran- 2-yl)methoxy)-2-(1H-tetrazol-5- yl)acetic acid 102

(R)-2-(((2R,3S,4R,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 3,4-dihydroxytetrahydrofuran- 2-yl)methoxy)-2-(1H-tetrazol-5- yl)acetic acid 103

(S)-2-(((2R,3S,4R,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 3,4-dihydroxytetrahydrofuran- 2-yl)methoxy)-3-phenyl-2-(1H- tetrazol-5-yl)propanic acid 104

(R)-2-(((2R,3S,4R,5R)-5-(6- amino-2-chloro-9H-purin-9-yl)- 3,4-dihydroxytetrahydrofuran- 2-yl)methoxy)-3-phenyl-2-(1H- tetrazol-5-yl)propanoic acid 105

(S)-2-(((2R,3S,4R,5R)-5-(6- amino-2-methoxy-9H-purin-9- yl)-3,4- dihydroxytetrahydrofuran-2- yl)methoxy)-3-phenyl-2-(1H- tetrazol-5-yl)propanic acid 106

(R)-2-(((2R,3S,4R,5R)-5-(6- amino-2-methoxy-9H-purin-9- yl)-3,4- dihydroxytetrahydrofuran-2- yl)methoxy)-3-phenyl-2-(1H- tetrazol-5-yl)propanic acid 107

Representative compounds of Formula (I) include those in Table 1B:

TABLE 1B Cpd. # Compound 108

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Other compounds of the invention include:

or a pharmaceutically acceptable salt thereof.

Pharmaceutical Preparations

In certain embodiments, the present invention provides a pharmaceutical preparation suitable for use in a human patient, comprising any of the compounds shown above (e.g., a compound of the invention, such as a compound of formula (I), and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical preparations may be for use in treating or preventing a condition or disease as described herein.

Any of the disclosed compounds may be used in the manufacture of medicaments for the treatment of any diseases or conditions disclosed herein.

Methods of Use Provided herein are methods of inhibiting CD73 in a cell, comprising contacting the cell with a compound of the invention, such as a compound of formula (I), or a pharmaceutically acceptable salt thereof. In certain embodiments, contacting the cell occurs in a subject in need thereof, thereby treating a disease or disorder mediated by adenosine.

Also, disclosed herein are methods of treating a disease or a disorder mediated by adenosine comprising administering a compound the invention, such as a compound of Formula (I), or a pharmaceutically acceptable salt thereof.

Adenosine acts on a variety of immune cells to induce immunosuppression, and the immunosuppressive effects of ectonucleotidases that enhance adenosine levels are also associated with enhanced infections of mammalian cells by parasites, fungi, bacteria, and viruses. Apart from immunosuppressive effects, adenosine also has a role in modulating the cardiovascular system (as a vasodilator and cardiac depressor), the central nervous system (CNS) (inducing sedative, anxiolytic and antiepileptic effects), the respiratory system (inducing bronchoconstriction), the kidney (having biphasic action; inducing vasoconstriction at low concentrations and vasodilation at high doses), fat cells (inhibiting lipolysis), and platelets (as an anti-aggregant). Furthermore, adenosine also promotes fibrosis (excess matrix production) in a variety of tissues. Therefore, improved treatments targeting CD73 would provide therapies for treating a wide range of conditions in addition to cancer, including cerebral and cardiac ischemic disease, fibrosis, immune and inflammatory disorders (e.g., inflammatory gut motility disorder), neurological, neurodegenerative and CNS disorders and diseases (e.g., depression, Parkinson's disease), and sleep disorders.

In some embodiments, the disease or the disorder mediated by adenosine is selected from cerebral ischemic disease, cancer, cardiac ischemic disease, depression, fibrosis, an immune disorder, an inflammatory disorder (e.g., inflammatory gut motility disorder), neurological disorder or disease, neurodegenerative disorder or disease (e.g., Parkinson's disease), CNS disorders and diseases, and sleep disorders.

The methods described herein are useful for the treatment of a wide variety of cancers, including bladder cancer, bone cancer, brain cancer (including glioblastoma), breast cancer, cardiac cancer, cervical cancer, colon cancer, colorectal cancer, esophageal cancer, fibrosarcoma, gastric cancer, gastrointestinal cancer, head & neck cancer, Kaposi's sarcoma, kidney cancer (including renal cell adenocarcinoma), leukemia, liver cancer, lung cancer (including non-small cell lung cancer, small cell lung cancer, and mucoepidermoid pulmonary carcinoma), lymphoma, melanoma, myeloma, ovarian cancer (including ovarian adenocarcinoma), pancreatic cancer, penile cancer, prostate cancer, testicular germcell cancer, thymoma and thymic carcinoma.

In some embodiments, the subject has a cancer selected from breast cancer, brain cancer, colon cancer, fibrosarcoma, kidney cancer, lung cancer, melanoma, ovarian cancer, and prostate cancer. In certain embodiments, the subject has a cancer selected from breast cancer, colon cancer, fibrosarcoma, melanoma, ovarian cancer, and prostate cancer. In other embodiments, the subject has a cancer selected from brain cancer, breast cancer, kidney cancer, lung cancer, melanoma, and ovarian cancer. In yet other embodiments, the subject has breast cancer. In some embodiments, the breast cancer is breast adenocarcinoma. In certain embodiments, the breast cancer is triple-negative breast cancer.

In certain embodiments, the methods for treating or preventing cancer can be demonstrated by one or more responses such as increased apoptosis, inhibition of tumor growth, reduction of tumor metastasis, inhibition of tumor metastasis, reduction of microvessel density, decreased neovascularization, inhibition of tumor migration, tumor regression, and increased survival of the subject.

In certain embodiments, the disease or the disorder mediated by adenosine is a disease or disorder mediated by CD73 activity. In some embodiments, the compounds of the invention, such as compounds of Formula (I), are useful as inhibitors of CD73.

In some embodiments, the methods described herein treat or prevent cardiovascular disease using inhibitors of CD73. Mutant genes encoding CD73 lead to extensive calcification of lower-extremity arteries and small joint capsules, which is associated with increased risk of cardiovascular disease (Hilaire et al., N. Engl. J. Med., 364(5): 432-442, 2011).

In some embodiments, the methods disclosed herein treat or prevent cancer using inhibitors of CD73. A CD73 small interfering RNA and anti-CD73 monoclonal antibodies showed a significant effect in treating or preventing cancer (Antonioli et al., Nat. Rev. Cancer, 13: 842-857, 2013). A tight correlation exists between CD73 expression and the ability of cancer cells to migrate, invade, and adhere to the extracellular matrix (ECM) (Antonioli 2013; Antonioli et al., Trends Cancer, 2(2): 95-109, 2016).

In some embodiments, the treatment or prevention of cancer by inhibitors of CD73 can be demonstrated by one or more responses selected from activation, clonal expansion, and homing of tumor-specific T cells (Antonioli 2016). In other embodiments, the methods disclosed herein increase the number of effector T lymphocytes (e.g., cytolytic effector T lymphocytes).

Combination Treatments

In some embodiments, the method of treating or preventing cancer may comprise administering a CD39 inhibitor conjointly with one or more other chemotherapeutic agent(s).

In one embodiment, the CD73 inhibitor is a compound of the invention, such as a compound of Formula (I). Other chemotherapeutic agents can include CD73-specific monoclonal antibodies which enhance the effects of other antibodies and therapies because of increased overall immune system activity (lower T-regulatory function and higher T-effector function, etc.) (Antonioli 2016).

In certain embodiments, the method of treating or preventing cancer may comprise administering a compound of the invention conjointly with one or more other chemotherapeutic agent(s).

Chemotherapeutic agents that may be conjointly administered with compounds of the invention include: 1-amino-4-phenylamino-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (acid blue 25), 1-amino-4-[4-hydroxyphenyl-amino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-aminophenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[1-naphthylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-fluoro-2-carboxyphenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[2-anthracenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, ABT-263, afatinib dimaleate, axitinib, aminoglutethimide, amsacrine, anastrozole, APCP, asparaginase, AZD5363, Bacillus Calmette-Guerin vaccine (bcg), bicalutamide, bleomycin, bortezomib, β-methylene-ADP (AOPCP), buserelin, busulfan, cabazitaxel, cabozantinib, campothecin, capecitabine, carboplatin, carfilzomib, carmustine, ceritinib, chlorambucil, chloroquine, cisplatin, cladribine, clodronate, cobimetinib, colchicine, crizotinib, cyclophosphamide, cyproterone, cytarabine, dacarbazine, dactinomycin, daunorubicin, demethoxyviridin, dexamethasone, dichloroacetate, dienestrol, diethylstilbestrol, docetaxel, doxorubicin, epirubicin, eribulin, erlotinib, estradiol, estramustine, etoposide, everolimus, exemestane, filgrastim, fludarabine, fludrocortisone, fluorouracil, fluoxymesterone, flutamide, gefitinib, gemcitabine, genistein, goserelin, GSK1120212, hydroxyurea, idarubicin, ifosfamide, imatinib, interferon, irinotecan, ixabepilone, lenalidomide, letrozole, leucovorin, leuprolide, levamisole, lomustine, lonidamine, mechlorethamine, medroxyprogesterone, megestrol, melphalan, mercaptopurine, mesna, metformin, methotrexate, miltefosine, mitomycin, mitotane, mitoxantrone, MK-2206, mutamycin, N-(4-sulfamoylphenylcarbamothioyl) pivalamide, NF279, NF449, nilutamide, nocodazole, octreotide, olaparib, oxaliplatin, paclitaxel, pamidronate, pazopanib, pemexetred, pentostatin, perifosine, PF-04691502, plicamycin, pomalidomide, porfimer, PPADS, procarbazine, quercetin, raltitrexed, ramucirumab, reactive blue 2, rituximab, rolofylline, romidepsin, rucaparib, selumetinib, sirolimus, sodium 2,4-dinitrobenzenesulfonate, sorafenib, streptozocin, sunitinib, suramin, talazoparib, tamoxifen, temozolomide, temsirolimus, teniposide, testosterone, thalidomide, thioguanine, thiotepa, titanocene dichloride, tonapofylline, topotecan, trametinib, trastuzumab, tretinoin, veliparib, vinblastine, vincristine, vindesine, vinorelbine, and vorinostat (SAHA). In other embodiments, chemotherapeutic agents that may be conjointly administered with compounds of the invention include: ABT-263, dexamethasone, 5-fluorouracil, PF-04691502, romidepsin, and vorinostat (SAHA). In other embodiments, chemotherapeutic agents that may be conjointly administered with compounds of the invention include: 1-amino-4-phenylamino-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (acid blue 25), 1-amino-4-[4-hydroxyphenyl-amino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-aminophenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[1-naphthylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-fluoro-2-carboxyphenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[2-anthracenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, APCP, β-methylene-ADP (AOPCP), capecitabine, cladribine, cytarabine, fludarabine, doxorubicin, gemcitabine, N-(4-sulfamoylphenylcarbamothioyl) pivalamide, NF279, NF449, PPADS, quercetin, reactive blue 2, rolofylline sodium 2,4-dinitrobenzenesulfonate, sumarin, and tonapofylline.

Many combination therapies have been developed for the treatment of cancer. In certain embodiments, compounds of the invention (e.g., compounds of Formula (I)) may be conjointly administered with a combination therapy. Examples of combination therapies with which compounds of the invention may be conjointly administered are included in Table 1.

TABLE 1 Exemplary combinatorial therapies for the treatment of cancer Name Therapeutic agents ABV Doxorubicin, Bleomycin, Vinblastine ABVD Doxorubicin, Bleomycin, Vinblastine, Dacarbazine AC (Breast) Doxorubicin, Cyclophosphamide AC Doxorubicin, Cisplatin (Sarcoma) AC (Neuro- Cyclophosphamide, Doxorubicin blastoma) ACE Cyclophosphamide, Doxorubicin, Etoposide ACe Cyclophosphamide, Doxorubicin AD Doxorubicin, Dacarbazine AP Doxorubicin, Cisplatin ARAC-DNR Cytarabine, Daunorubicin B-CAVe Bleomycin, Lomustine, Doxorubicin, Vinblastine BCVPP Carmustine, Cyclophosphamide, Vinblastine, Procarbazine, Prednisone BEACOPP Bleomycin, Etoposide, Doxorubicin, Cyclophosphamide, Vincristine, Procarbazine, Prednisone, Filgrastim BEP Bleomycin, Etoposide, Cisplatin BIP Bleomycin, Cisplatin, Ifosfamide, Mesna BOMP Bleomycin, Vincristine, Cisplatin, Mitomycin CA Cytarabine, Asparaginase CABO Cisplatin, Methotrexate, Bleomycin, Vincristine CAF Cyclophosphamide, Doxorubicin, Fluorouracil CAL-G Cyclophosphamide, Daunorubicin, Vincristine, Prednisone, Asparaginase CAMP Cyclophosphamide, Doxorubicin, Methotrexate, Procarbazine CAP Cyclophosphamide, Doxorubicin, Cisplatin CAV Cyclophosphamide, Doxorubicin, Vincristine CAVE ADD CAV and Etoposide CA-VP16 Cyclophosphamide, Doxorubicin, Etoposide CC Cyclophosphamide, Carboplatin CDDP/ Cisplatin, Etoposide VP-16 CEF Cyclophosphamide, Epirubicin, Fluorouracil CEPP(B) Cyclophosphamide, Etoposide, Prednisone, with or without/Bleomycin CEV Cyclophosphamide, Etoposide, Vincristine CF Cisplatin, Fluorouracil or Carboplatin Fluorouracil CHAP Cyclophosphamide or Cyclophosphamide, Altretamine, Doxorubicin, Cisplatin ChlVPP Chlorambucil, Vinblastine, Procarbazine, Prednisone CHOP Cyclophosphamide, Doxorubicin, Vincristine, Prednisone CHOP- Add Bleomycin to CHOP BLEO CISCA Cyclophosphamide, Doxorubicin, Cisplatin CLD-BOMP Bleomycin, Cisplatin, Vincristine, Mitomycin CMF Methotrexate, Fluorouracil, Cyclophosphamide CMFP Cyclophosphamide, Methotrexate, Fluorouracil, Prednisone CMFVP Cyclophosphamide, Methotrexate, Fluorouracil, Vincristine, Prednisone CMV Cisplatin, Methotrexate, Vinblastine CNF Cyclophosphamide, Mitoxantrone, Fluorouracil CNOP Cyclophosphamide, Mitoxantrone, Vincristine, Prednisone COB Cisplatin, Vincristine, Bleomycin CODE Cisplatin, Vincristine, Doxorubicin, Etoposide COMLA Cyclophosphamide, Vincristine, Methotrexate, Leucovorin, Cytarabine COMP Cyclophosphamide, Vincristine, Methotrexate, Prednisone Cooper Cyclophosphamide, Methotrexate, Fluorouracil, Regimen Vincristine, Prednisone COP Cyclophosphamide, Vincristine, Prednisone COPE Cyclophosphamide, Vincristine, Cisplatin, Etoposide COPP Cyclophosphamide, Vincristine, Procarbazine, Prednisone CP(Chronic Chlorambucil, Prednisone lymphocytic leukemia) CP (Ovarian Cyclophosphamide, Cisplatin Cancer) CVD Cisplatin, Vinblastine, Dacarbazine CVI Carboplatin, Etoposide, Ifosfamide, Mesna CVP Cyclophosphamide, Vincristine, Prednisome CVPP Lomustine, Procarbazine, Prednisone CYVADIC Cyclophosphamide, Vincristine, Doxorubicin, Dacarbazine DA Daunorubicin, Cytarabine DAT Daunorubicin, Cytarabine, Thioguanine DAV Daunorubicin, Cytarabine, Etoposide DCT Daunorubicin, Cytarabine, Thioguanine DHAP Cisplatin, Cytarabine, Dexamethasone DI Doxorubicin, Ifosfamide DTIC/ Dacarbazine, Tamoxifen Tamoxifen DVP Daunorubicin, Vincristine, Prednisone EAP Etoposide, Doxorubicin, Cisplatin EC Etoposide, Carboplatin EFP Etoposie, Fluorouracil, Cisplatin ELF Etoposide, Leucovorin, Fluorouracil EMA 86 Mitoxantrone, Etoposide, Cytarabine EP Etoposide, Cisplatin EVA Etoposide, Vinblastine FAC Fluorouracil, Doxorubicin, Cyclophosphamide FAM Fluorouracil, Doxorubicin, Mitomycin FAMTX Methotrexate, Leucovorin, Doxorubicin FAP Fluorouracil, Doxorubicin, Cisplatin F-CL Fluorouracil, Leucovorin FEC Fluorouracil, Cyclophosphamide, Epirubicin FED Fluorouracil, Etoposide, Cisplatin FL Flutamide, Leuprolide FZ Flutamide, Goserelin acetate implant HDMTX Methotrexate, Leucovorin Hexa-CAF Altretamine, Cyclophosphamide, Methotrexate, Fluorouracil IDMTX/ Methotrexate, Mercaptopurine, Leucovorin 6-MP IE Ifosfamide, Etoposie, Mesna IfoVP Ifosfamide, Etoposide, Mesna IPA Ifosfamide, Cisplatin, Doxorubicin M-2 Vincristine, Carmustine, Cyclophosphamide, Prednisone, Melphalan MAC-III Methotrexate, Leucovorin, Dactinomycin, Cyclophosphamide MACC Methotrexate, Doxorubicin, Cyclophosphamide, Lomustine MACOP-B Methotrexate, Leucovorin, Doxorubicin, Cyclophosphamide, Vincristine, Bleomycin, Prednisone MAID Mesna, Doxorubicin, Ifosfamide, Dacarbazine m-BACOD Bleomycin, Doxorubicin, Cyclophosphamide, Vincristine, Dexamethasone, Methotrexate, Leucovorin MBC Methotrexate, Bleomycin, Cisplatin MC Mitoxantrone, Cytarabine MF Methotrexate, Fluorouracil, Leucovorin MICE Ifosfamide, Carboplatin, Etoposide, Mesna MINE Mesna, Ifosfamide, Mitoxantrone, Etoposide mini-BEAM Carmustine, Etoposide, Cytarabine, Melphalan MOBP Bleomycin, Vincristine, Cisplatin, Mitomycin MOP Mechlorethamine, Vincristine, Procarbazine MOPP Mechlorethamine, Vincristine, Procarbazine, Prednisone MOPP/ABV Mechlorethamine, Vincristine, Procarbazine, Prednisone, Doxorubicin, Bleomycin, Vinblastine MP (multiple Melphalan, Prednisone myeloma) MP (prostate Mitoxantrone, Prednisone cancer) MTX/6-MO Methotrexate, Mercaptopurine MTX/6- Methotrexate, Mercaptopurine, Vincristine, Prednisone MP/VP MTX- Methotrexate, Leucovorin, Cisplatin, Doxorubicin CDDPAdr MV (breast Mitomycin, Vinblastine cancer) MV (acute Mitoxantrone, Etoposide myelocytic leukemia) M-VAC Vinblastine, Doxorubicin, Cisplatin Methotrexate MVP Vinblastine, Cisplatin Mitomycin MVPP Mechlorethamine, Vinblastine, Procarbazine, Prednisone NFL Mitoxantrone, Fluorouracil, Leucovorin NOVP Mitoxantrone, Vinblastine, Vincristine OPA Vincristine, Prednisone, Doxorubicin OPPA Add Procarbazine to OPA. PAC Cisplatin, Doxorubicin PAC-I Cisplatin, Doxorubicin, Cyclophosphamide PA-CI Cisplatin, Doxorubicin PCV Lomustine, Procarbazine, Vincristine PFL Cisplatin, Fluorouracil, Leucovorin POC Prednisone, Vincristine, Lomustine ProMACE Prednisone, Methotrexate, Leucovorin, Doxorubicin, Cyclophosphamide, Etoposide ProMACE/ Prednisone, Doxorubicin, Cyclophosphamide, Etoposide, cytaBOM Cytarabine, Bleomycin, Vincristine, Methotrexate, Leucovorin, Cotrimoxazole PRoMACE/ Prednisone, Doxorubicin, Cyclophosphamide, Etoposide, MOPP Mechlorethamine, Vincristine, Procarbazine, Methotrexate, Leucovorin Pt/VM Cisplatin, Teniposide PVA Prednisone, Vincristine, Asparaginase PVB Cisplatin, Vinblastine, Bleomycin PVDA Prednisone, Vincristine, Daunorubicin, Asparaginase SMF Streptozocin, Mitomycin, Fluorouracil TAD Mechlorethamine, Doxorubicin, Vinblastine, Vincristine, Bleomycin, Etoposide, Prednisone TTT Methotrexate, Cytarabine, Hydrocortisone Topo/CTX Cyclophosphamide, Topotecan, Mesna VAB-6 Cyclophosphamide, Dactinomycin, Vinblastine, Cisplatin, Bleomycin VAC Vincristine, Dactinomycin, Cyclophosphamide VACAdr Vincristine, Cyclophosphamide, Doxorubicin, Dactinomycin, Vincristine VAD Vincristine, Doxorubicin, Dexamethasone VATH Vinblastine, Doxorubicin, Thiotepa, Flouxymesterone VBAP Vincristine, Carmustine, Doxorubicin, Prednisone VBCMP Vincristine, Carmustine, Melphalan, Cyclophosphamide, Prednisone VC Vinorelbine, Cisplatin VCAP Vincristine, Cyclophosphamide, Doxorubicin, Prednisone VD Vinorelbine, Doxorubicin VelP Vinblastine, Cisplatin, Ifosfamide, Mesna VIP Etoposide, Cisplatin, Ifosfamide, Mesna VM Mitomycin, Vinblastine VMCP Vincristine, Melphalan, Cyclophosphamide, Prednisone VP Etoposide, Cisplatin V-TAD Etoposide, Thioguanine, Daunorubicin, Cytarabine 5 + 2 Cytarabine, Daunorubicin, Mitoxantrone 7 + 3 Cytarabine with/, Daunorubicin or Idarubicin or Mitoxantrone “8 in 1” Methylprednisolone, Vincristine, Lomustine, Procarbazine, Hydroxyurea, Cisplatin, Cytarabine, Dacarbazine

In some embodiments, the chemotherapeutic agents that may be conjointly administered with compounds of the invention, such as a compound of Formula (I), include a CD39 inhibitor. CD39 or ecto-nucleoside triphosphate diphosphohydrolase 1 (E-NTPDase1 or ENTPD 1) is a membrane-bound enzyme that catalyzes the conversion of extracellular adenosine triphosphate (ATP) and/or ADP (adenosine diphosphate) to adenosine monophosphate (AMP). In one embodiment, the CD39 inhibitor is polyoxometalate-1 (POM-1).

In other embodiments, the chemotherapeutic agents that may be conjointly administered with compounds of the invention, such as a compound of Formula (I), include known CD73 inhibitors. In some embodiments, the CD73 inhibitor is an anthraquinone derivative (Baqi et al., J. Med. Chem., 53(5): 2076-2086, 2010, herein incorporated by reference). In other embodiments, the CD73 inhibitor is an sulfonic acid derivative (Raza et al., Med. Chem., 8: 1133-1139, 2012, herein incorporated by reference). In yet other embodiments, the CD73 inhibitor is selected from 1-amino-4-phenylamino-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate (acid blue 25), 1-amino-4-[4-hydroxyphenyl-amino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-aminophenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[1-naphthylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[4-fluoro-2-carboxyphenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, 1-amino-4-[2-anthracenylamino]-9,10-dioxo-9,10-dihydroanthracene-2-sulfonate, sodium 2,4-dinitrobenzenesulfonate, N-(4-sulfamoylphenylcarbamothioyl) pivalamide, APCP, β-methylene-ADP (AOPCP), PPADS, NF279, NF449, quercetin, reactive blue 2, and sumarin (Baqi 2010; Raza 2012).

In certain embodiments, the combination of a compound of the invention, such as a compound of Formula (I), with a second CD73 inhibitor or a CD39 inhibitor may have a synergistic effect in the treatment of cancer and other diseases or disorders mediated by adenosine. Without wishing to be bound by any theory, this synergy may be observed because CD39 and CD73 are often on different cell types. The hypoxic tumor microenvironment also induces greater levels of CD39 and CD73.

In some embodiments, the chemotherapeutic agents that may be conjointly administered with compounds of the invention, such as a compound of Formula (I), include an adenosine receptor inhibitor. In other embodiments, the adenosine receptor inhibitor is selected from rolofylline, tonapofylline, ATL-444, istradefylline, MSX-3, preladenant, SCH-58,261, SCH-412,348, SCH-442,416, ST-1535, VER-6623, VER-6947, VER-7835, vipadenant, and ZM-241,385. In some embodiments, the adenosine receptor inhibitor targets the A_(2A) receptor as this subtype is predominantly expressed in most immune cells.

In other embodiments, the chemotherapeutic agents that may be conjointly administered with compounds of the invention, such as a compound of Formula (I), include a nucleoside-based drug. In certain embodiments, the nucleoside-based drug is selected from gemcitabine, capecitabine, cytarabine, fludarabine and cladribine.

In further embodiments, the combination therapy comprises a compound of the invention, such as a compound of Formula (I), conjointly administered with an anthracycline. In other embodiments, the combination therapy comprises a compound of the invention, such as a compound of Formula (I), conjointly administered with doxorubicin. Combination treatment with an anti-CD73 antibody and doxorubicin has demonstrated a significant chemotherapeutic effect (Young et al., Cancer Discov., 4(8): 1-10, 2014, herein incorporated by reference).

In certain embodiments, the combination therapy comprises a compound of the invention, such as a compound of Formula (I), conjointly administered with an A_(2A) receptor inhibitor and an anthracycline. In some embodiments, the anthracycline is doxorubicin. Combination treatment with an anti-CD73 antibody, an A_(2A) receptor inhibitor, and doxorubicin has demonstrated an increased chemotherapeutic effect (Antonioli 2013).

In certain embodiments, the conjoint therapies of the invention comprise conjoint administration with other types of chemotherapeutic agents, such as immuno-oncology agents. Cancer cells often have specific cell surface antigens that can be recognized by the immune system. Thus, immuno-oncology agents, such as monoclonal antibodies, can selectively bind to cancer cell antigens and effect cell death. Other immuno-oncology agents can suppress tumor-mediated inhibition of the native immune response or otherwise activate the immune response and thus facilitate recognition of the tumor by the immune system. Exemplary antibody immuno-oncology agents, include, but are not limited to, abagovomab, adecatumumab, afutuzumab, alemtuzumab, anatumomab mafenatox, apolizumab, blinatumomab, BMS-936559, catumaxomab, durvalumab, epacadostat, epratuzumab, indoximod, inotuzumab ozogamicin, intelumumab, ipilimumab, isatuximab, lambrolizumab, MED14736, MPDL3280A, nivolumab, obinutuzumab, ocaratuzumab, ofatumumab, olatatumab, pembrolizumab, pidilizumab, rituximab, ticilimumab, samalizumab, and tremelimumab. In some embodiments, the antibody immune-oncology agents are selected from anti-CD73 monoclonal antibody (mAb), anti-CD39 mAb, anti-PD-1 mAb, and anti-CTLA4 mAb. Thus, in some embodiments, the methods of the invention comprise conjoint administration of one or more immuno-oncology agents, such as the agents mentioned above.

In some embodiments, the combination therapy comprises a compound of the invention, such as a compound of Formula (I), conjointly administered with anti-PD-1 therapy and anti-CTLA4 therapy. Combination treatment with an anti-CD73 monoclonal antibody (mAb), anti-PD-1 mAb, and anti-CTLA4 mAb showed a significant chemotherapeutic effect (Young 2014; Antonioli 2013).

In certain embodiments, a compound of the invention may be conjointly administered with non-chemical methods of cancer treatment. In certain embodiments, a compound of the invention may be conjointly administered with radiation therapy. In certain embodiments, a compound of the invention may be conjointly administered with surgery, with thermoablation, with focused ultrasound therapy, with cryotherapy, or with any combination of these.

In certain embodiments, compounds of the invention may be conjointly administered with one or more other compounds of the invention. Moreover, such combinations may be conjointly administered with other therapeutic agents, such as other agents suitable for the treatment of cancer, immunological or neurological diseases, such as the agents identified above. In certain embodiments, conjointly administering one or more additional chemotherapeutic agents with a compound of the invention provides a synergistic effect. In certain embodiments, conjointly administering one or more additional chemotherapeutic agents provides an additive effect.

Pharmaceutical Compositions

The compositions and methods of the present invention may be utilized to treat a subject in need thereof. In certain embodiments, the subject is a mammal such as a human, or a non-human mammal. When administered to subject, such as a human, the composition or the compound is preferably administered as a pharmaceutical composition comprising, for example, a compound of the invention and a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are well known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or vehicles such as glycols, glycerol, oils such as olive oil, or injectable organic esters. In a preferred embodiment, when such pharmaceutical compositions are for human administration, particularly for invasive routes of administration (i.e., routes, such as injection or implantation, that circumvent transport or diffusion through an epithelial barrier), the aqueous solution is pyrogen-free, or substantially pyrogen-free. The excipients can be chosen, for example, to effect delayed release of an agent or to selectively target one or more cells, tissues or organs. The pharmaceutical composition can be in dosage unit form such as tablet, capsule (including sprinkle capsule and gelatin capsule), granule, lyophile for reconstitution, powder, solution, syrup, suppository, injection or the like. The composition can also be present in a transdermal delivery system, e.g., a skin patch. The composition can also be present in a solution suitable for topical administration, such as an eye drop.

A pharmaceutically acceptable carrier can contain physiologically acceptable agents that act, for example, to stabilize, increase solubility or to increase the absorption of a compound such as a compound of the invention. Such physiologically acceptable agents include, for example, carbohydrates, such as glucose, sucrose or dextrans, antioxidants, such as ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The choice of a pharmaceutically acceptable carrier, including a physiologically acceptable agent, depends, for example, on the route of administration of the composition. The preparation or pharmaceutical composition can be a self-emulsifying drug delivery system or a self-microemulsifying drug delivery system. The pharmaceutical composition (preparation) also can be a liposome or other polymer matrix, which can have incorporated therein, for example, a compound of the invention. Liposomes, for example, which comprise phospholipids or other lipids, are nontoxic, physiologically acceptable and metabolizable carriers that are relatively simple to make and administer.

The phrase “pharmaceutically acceptable” is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of a subject without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

The phrase “pharmaceutically acceptable carrier” as used herein means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the subject. Some examples of materials which can serve as pharmaceutically acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as corn starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) phosphate buffer solutions; and (21) other non-toxic compatible substances employed in pharmaceutical formulations.

A pharmaceutical composition (preparation) can be administered to a subject by any of a number of routes of administration including, for example, orally (for example, drenches as in aqueous or non-aqueous solutions or suspensions, tablets, capsules (including sprinkle capsules and gelatin capsules), boluses, powders, granules, pastes for application to the tongue); absorption through the oral mucosa (e.g., sublingually); anally, rectally or vaginally (for example, as a pessary, cream or foam); parenterally (including intramuscularly, intravenously, subcutaneously or intrathecally as, for example, a sterile solution or suspension); nasally; intraperitoneally; subcutaneously; transdermally (for example as a patch applied to the skin); and topically (for example, as a cream, ointment or spray applied to the skin, or as an eye drop). The compound may also be formulated for inhalation. In certain embodiments, a compound may be simply dissolved or suspended in sterile water. Details of appropriate routes of administration and compositions suitable for same can be found in, for example, U.S. Pat. Nos. 6,110,973, 5,763,493, 5,731,000, 5,541,231, 5,427,798, 5,358,970 and 4,172,896, as well as in patents cited therein.

The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the subject being treated, the particular mode of administration. The amount of active ingredient that can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. Generally, out of one hundred percent, this amount will range from about 1 percent to about ninety-nine percent of active ingredient, preferably from about 5 percent to about 70 percent, most preferably from about 10 percent to about 30 percent.

Methods of preparing these formulations or compositions include the step of bringing into association an active compound, such as a compound of the invention, with the carrier and, optionally, one or more accessory ingredients. In general, the formulations are prepared by uniformly and intimately bringing into association a compound of the present invention with liquid carriers, or finely divided solid carriers, or both, and then, if necessary, shaping the product.

Formulations of the invention suitable for oral administration may be in the form of capsules (including sprinkle capsules and gelatin capsules), cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), lyophile, powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and/or as mouth washes and the like, each containing a predetermined amount of a compound of the present invention as an active ingredient. Compositions or compounds may also be administered as a bolus, electuary or paste.

To prepare solid dosage forms for oral administration (capsules (including sprinkle capsules and gelatin capsules), tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically acceptable carriers, such as sodium citrate or dicalcium phosphate, and/or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and/or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof, (10) complexing agents, such as, modified and unmodified cyclodextrins; and (11) coloring agents. In the case of capsules (including sprinkle capsules and gelatin capsules), tablets and pills, the pharmaceutical compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.

A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active or dispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceutical compositions, such as dragees, capsules (including sprinkle capsules and gelatin capsules), pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and/or microspheres. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the above-described excipients.

Liquid dosage forms useful for oral administration include pharmaceutically acceptable emulsions, lyophiles for reconstitution, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, cyclodextrins and derivatives thereof, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.

Formulations of the pharmaceutical compositions for rectal, vaginal, or urethral administration may be presented as a suppository, which may be prepared by mixing one or more active compounds with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.

Formulations of the pharmaceutical compositions for administration to the mouth may be presented as a mouthwash, or an oral spray, or an oral ointment.

Alternatively or additionally, compositions can be formulated for delivery via a catheter, stent, wire, or other intraluminal device. Delivery via such devices may be especially useful for delivery to the bladder, urethra, ureter, rectum, or intestine.

Formulations which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.

Dosage forms for the topical or transdermal administration include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically acceptable carrier, and with any preservatives, buffers, or propellants that may be required.

The ointments, pastes, creams and gels may contain, in addition to an active compound, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to an active compound, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

Transdermal patches have the added advantage of providing controlled delivery of a compound of the present invention to the body. Such dosage forms can be made by dissolving or dispersing the active compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like, are also contemplated as being within the scope of this invention. Exemplary ophthalmic formulations are described in U.S. Publication Nos. 2005/0080056, 2005/0059744, 2005/0031697 and 2005/004074 and U.S. Pat. No. 6,583,124, the contents of which are incorporated herein by reference. If desired, liquid ophthalmic formulations have properties similar to that of lacrimal fluids, aqueous humor or vitreous humor or are compatible with such fluids. A preferred route of administration is local administration (e.g., topical administration, such as eye drops, or administration via an implant).

The phrases “parenteral administration” and “administered parenterally” as used herein means modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

Pharmaceutical compositions suitable for parenteral administration comprise one or more active compounds in combination with one or more pharmaceutically acceptable sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.

These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents, for example, paraben, chlorobutanol, phenol sorbic acid, and the like. It may also be desirable to include isotonic agents, such as sugars, sodium chloride, and the like into the compositions. In addition, prolonged absorption of the injectable pharmaceutical form may be brought about by the inclusion of agents that delay absorption such as aluminum monostearate and gelatin.

In some cases, in order to prolong the effect of a drug, it is desirable to slow the absorption of the drug from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material having poor water solubility. The rate of absorption of the drug then depends upon its rate of dissolution, which, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered drug form is accomplished by dissolving or suspending the drug in an oil vehicle.

Injectable depot forms are made by forming microencapsulated matrices of the subject compounds in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of drug to polymer, and the nature of the particular polymer employed, the rate of drug release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping the drug in liposomes or microemulsions that are compatible with body tissue.

For use in the methods of this invention, active compounds can be given per se or as a pharmaceutical composition containing, for example, 0.1 to 99.5% (more preferably, 0.5 to 90%) of active ingredient in combination with a pharmaceutically acceptable carrier.

Methods of introduction may also be provided by rechargeable or biodegradable devices. Various slow release polymeric devices have been developed and tested in vivo in recent years for the controlled delivery of drugs, including proteinacious biopharmaceuticals. A variety of biocompatible polymers (including hydrogels), including both biodegradable and non-degradable polymers, can be used to form an implant for the sustained release of a compound at a particular target site.

Actual dosage levels of the active ingredients in the pharmaceutical compositions may be varied so as to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factors including the activity of the particular compound or combination of compounds employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion of the particular compound(s) being employed, the duration of the treatment, other drugs, compounds and/or materials used in combination with the particular compound(s) employed, the age, sex, weight, condition, general health and prior medical history of the subject being treated, and like factors well known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the therapeutically effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could start doses of the pharmaceutical composition or compound at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. By “therapeutically effective amount” is meant the concentration of a compound that is sufficient to elicit the desired therapeutic effect. It is generally understood that the effective amount of the compound will vary according to the weight, sex, age, and medical history of the subject. Other factors which influence the effective amount may include, but are not limited to, the severity of the subject's condition, the disorder being treated, the stability of the compound, and, if desired, another type of therapeutic agent being administered with the compound of the invention. A larger total dose can be delivered by multiple administrations of the agent. Methods to determine efficacy and dosage are known to those skilled in the art (Isselbacher et al. (1996) Harrison's Principles of Internal Medicine 13 ed., 1814-1882, herein incorporated by reference).

In general, a suitable daily dose of an active compound used in the compositions and methods of the invention will be that amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.

If desired, the effective daily dose of the active compound may be administered as one, two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms. In certain embodiments of the present invention, the active compound may be administered two or three times daily. In preferred embodiments, the active compound will be administered once daily.

In certain embodiments, the dosing follows a 3+3 design. The traditional 3+3 design requires no modeling of the dose-toxicity curve beyond the classical assumption for cytotoxic drugs that toxicity increases with dose. This rule-based design proceeds with cohorts of three patients; the first cohort is treated at a starting dose that is considered to be safe based on extrapolation from animal toxicological data, and the subsequent cohorts are treated at increasing dose levels that have been fixed in advance. In some embodiments, the three doses of a compound of formula (I) range from about 100 mg to about 1000 mg orally, such as about 200 mg to about 800 mg, such as about 400 mg to about 700 mg, such as about 100 mg to about 400 mg, such as about 500 mg to about 1000 mg, and further such as about 500 mg to about 600 mg. Dosing can be three times a day when taken with without food, or twice a day when taken with food. In certain embodiments, the three doses of a compound of formula (I) range from about 400 mg to about 800 mg, such as about 400 mg to about 700 mg, such as about 500 mg to about 800 mg, and further such as about 500 mg to about 600 mg twice a day. In certain preferred embodiments, a dose of greater than about 600 mg is dosed twice a day.

If none of the three patients in a cohort experiences a dose-limiting toxicity, another three patients will be treated at the next higher dose level. However, if one of the first three patients experiences a dose-limiting toxicity, three more patients will be treated at the same dose level. The dose escalation continues until at least two patients among a cohort of three to six patients experience dose-limiting toxicities (i.e., ≥about 33% of patients with a dose-limiting toxicity at that dose level). The recommended dose for phase II trials is conventionally defined as the dose level just below this toxic dose level.

In certain embodiments, the dosing schedule can be about 40 mg/m² to about 100 mg/m², such as about 50 mg/m² to about 80 mg/m², and further such as about 70 mg/m² to about 90 mg/m² by IV for 3 weeks of a 4 week cycle.

In certain embodiments, compounds of the invention may be used alone or conjointly administered with another type of therapeutic agent. As used herein, the phrase “conjoint administration” refers to any form of administration of two or more different therapeutic compounds such that the second compound is administered while the previously administered therapeutic compound is still effective in the body (e.g., the two compounds are simultaneously effective in the subject, which may include synergistic effects of the two compounds). For example, the different therapeutic compounds can be administered either in the same formulation or in a separate formulation, either concomitantly or sequentially. In certain embodiments, the different therapeutic compounds can be administered within one hour, 12 hours, 24 hours, 36 hours, 48 hours, 72 hours, or a week of one another. Thus, a subject who receives such treatment can benefit from a combined effect of different therapeutic compounds.

In certain embodiments, conjoint administration of compounds of the invention with one or more additional therapeutic agent(s) (e.g., one or more additional chemotherapeutic agent(s)) provides improved efficacy relative to each individual administration of the compound of the invention (e.g., compound of formula I or Ia) or the one or more additional therapeutic agent(s). In certain such embodiments, the conjoint administration provides an additive effect, wherein an additive effect refers to the sum of each of the effects of individual administration of the compound of the invention and the one or more additional therapeutic agent(s).

This invention includes the use of pharmaceutically acceptable salts of compounds of the invention in the compositions and methods of the present invention. In certain embodiments, contemplated salts of the invention include, but are not limited to, alkyl, dialkyl, trialkyl or tetra-alkyl ammonium salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, L-arginine, benenthamine, benzathine, betaine, calcium hydroxide, choline, deanol, diethanolamine, diethylamine, 2-(diethylamino)ethanol, ethanolamine, ethylenediamine, N-methylglucamine, hydrabamine, 1H-imidazole, lithium, L-lysine, magnesium, 4-(2-hydroxyethyl)morpholine, piperazine, potassium, 1-(2-hydroxyethyl)pyrrolidine, sodium, triethanolamine, tromethamine, and zinc salts. In certain embodiments, contemplated salts of the invention include, but are not limited to, Na, Ca, K, Mg, Zn or other metal salts.

The pharmaceutically acceptable acid addition salts can also exist as various solvates, such as with water, methanol, ethanol, dimethylformamide, and the like. Mixtures of such solvates can also be prepared. The source of such solvate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent.

Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically acceptable antioxidants include: (1) water-soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal-chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

General Synthesis

Compound numbers 1-45 as used in the general synthesis section below refer only to genus structures in this section and do not apply to compounds disclosed elsewhere in this application. Compounds disclosed herein can be made by methods depicted in the reaction schemes below.

The starting materials and reagents used in preparing these compounds are either available from commercial supplier such as Aldrich Chemical Co., Bachem, etc., or can be made by methods well known in the art. The schemes are merely illustrative of some methods by which the compounds disclosed herein can be synthesized and various modifications to these schemes can be made and will be suggested to POSITA having referred to this disclosure. The starting materials and the intermediates and the final products of the reaction may be isolated and purified if desired using convential techniques, including but not limited to filtration, distillation, crystallization, chromatography, and the like and may be characterized using conventional means, including physical constants and spectral data.

Unless specified otherwise, the reactions described herein take place at atmospheric pressure over a temperature range from about −78° C. to about 150° C.

Compounds of Formula (I) having the structure:

where R^(u), R^(v), R⁵, and R⁹ are as defined in the Summary can be synthesized as illustrated and described in Scheme 1 below.

Removal of the benzoyl groups in a compound of formula 1 with a suitable base such as NH₃ in an organic alcohol solvent such as MeOH, or aq. LiOH or aq. NaOH, followed by selectively protecting the 5′-OH group with tert-butyldiphenylsilyl group provides a compound of formula 2.

Compounds of formula 1 are either commercially available or they can be prepared by methods well known in the art. For example, ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(2,6-dichloro-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methyl benzoate and ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(2-amino-6-chloro-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methyl benzoate are commercially available. Compounds of formula 1 where R^(u) is other than chloro or amino and R^(v) is other than chloro can be prepared from ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(2,6-dichloro-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methyl benzoate as described in PCT application publication nos. WO 2015/164573 and WO 2017/120508.

Protection of the 3′-hydroxy group in compound 2 with a suitable hydroxy protecting group such as Boc, followed by removal of the silyl group under conditions well known in the art such as TBAF in THF provides a compound of formula 4. Reaction of compound 4 with a diazo reagent of formula 5 where R⁹ is unsubstituted alkyl or benzyl in the presence of catalyst, such as Rh₂(OAc)₄ or CuOAc₂ in a suitable organic solvent such as toluene, benzene, dichloromethane, dichloroethane, and THF, provides a compound of Formula 6 where R⁵ is hydrogen. Compound 6 can then be converted to a compound of Formula (I) by removal of the Boc group under acidic hydrolysis conditions, followed by removal of the R⁹ group under basic hydrolysis reaction condition e.g, aq. LiOH or NaOH (when R⁹ is alkyl) or by hydrogenolysis with Pd/C or Pd(OH)₂/C (when R⁹ is benzyl).

Alternatively, compound 6 can be reacted with a halide of formula R⁵X where X is halo (preferably chloro, bromo, iodo, tosylate, mesylate or triflate) and R⁵ is as defined in the Summary except hydrogen under alkylating reaction conditions to provide a corresponding compound of Formula (I) where R⁵ and R⁹ are other than hydrogen which can then be converted to corresponding compound of Formula (I) where R⁹ are hydrogen as described above.

Compounds of Formula (I) where Het is other than purine can be prepared by methods well known in the art. For example, compounds of Formula (I) where is a group of formula (i), (ii), or (vii) can be prepared from 2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine, 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine and 4,6-dichloro-1H-imidazo[4,5-c]pyridine respectively, by synthetic procedures disclosed in PCT application publication no. WO 2017/120508 and Scheme 1 above.

Proceeding as described above but substituting compound of formula 1, with (2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-chloro-2-(hydroxymethyl)-tetrahydrofuran-3-ol (prepared according to the procedures reported by Secrist, John A., III et al, Journal of Medicinal Chemistry, 31(2), 405-10; 1988 and by Anderson, Bruce G. et al., Organic Process Research & Development, 12(6), 1229-1237; 2008), compounds of Formula (I) where R^(1a) is chloro, R^(1b) and R^(2b) are hydrogen, and R^(2a) is hydroxy and Het is 2-chloro-9H-purin-6-amine can be prepared. It will be apparent to a person skilled that analogs of such compounds i.e, Het is other than 2-chloro-9H-purin-6-amine can also be prepared based on the disclosure of this Application and methods known in the art.

Proceeding as described above but substituting compound of formula 1, with (2R,3S,4S,5R)-2-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-5-(hydroxymethyl)-tetrahydrofuran-3-ol (prepared according to procedures reported by Ren, Hang et al Beilstein Journal of Organic Chemistry, 11, 2509-2520; 2015, and by Schinazi, Raymond F. et al., Heterocyclic Communications, 21(5), 315-327; 2015) compounds of Formula (I) where R^(1a) is hydroxy, R^(1b) and R^(2b) are hydrogen, and R^(2a) is fluoro and Het is 2-chloro-9H-purin-6-amine can be prepared. It will be apparent to a person skilled that analogs of such compounds i.e, Het is other than 2-chloro-9H-purin-6-amine can also be prepared based on the disclosure of this Application and methods known in the art.

Proceeding as described above but substituting compound of formula 1, with ((2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-difluorotetrahydrofuran-2-yl)methanol (prepared according to procedures reported by Schinazi, Raymond F. et al., Heterocyclic Communications, 21(5), 315-327; 2015 and by Sivets, Grigorii G. et al., Nucleosides, Nucleotides & Nucleic Acids, 28(5-7), 519-536; 2009) compounds of Formula (I) where R^(1b) is fluoro, R^(1a) and R^(2a) are hydrogen, and R^(2b) is fluoro and Het is 2-chloro-9H-purin-6-amine can be prepared. It will be apparent to a person skilled that analogs of such compounds i.e, Het is other than 2-chloro-9H-purin-6-amine can also be prepared based on the disclosure of this Application and methods known in the art.

Proceeding as described above but substituting compound of formula 1, with (2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-2-(hydroxymethyl)-3-methyltetrahydrofuran-3,4-diol (prepared according to the procedures reported by Franchetti, Palmarisa, et al., J. Med. Chem., 48(15), 4983-4989, 2005), or (2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3-ethynyl-2-(hydroxymethyl)tetrahydrofuran-3,4-diol (prepared according to the procedures reported by Hulpia, Fabian et al., Bioorganic & Medicinal Chemistry Letters, 26(8), 1970-1972; 2016) compounds of Formula (I) where R^(1a) and R^(2a) are hydroxy, R^(1b) is hydrogen, and R^(2b) is methyl or ethynyl respectively, and Het is 2-chloro-9H-purin-6-amine can be prepared. It will be apparent to a person skilled that analogs of such compounds i.e, Het is other than 2-chloro-9H-purin-6-amine can also be prepared based on the disclosure of this Application and methods known in the art.

Compounds of Formula (I) having the structure:

where R^(1a) is halo, alkyl, alkynyl, alkenyl, or cyano, R^(1b) is halo or hydroxy, or R^(1a) and R^(1b) are fluoro, Het is a ring of formula (iii) where R^(s) is hydrogen, and R^(u) and R^(v) are as defined in the Summary and R⁵ and R⁹ are as defined in the Summary can be synthesized by proceeding as described in Scheme 2 below.

Compounds of Formula (I) where R^(1a) is halo, alkyl, alkynyl, alkenyl, or cyano, R^(1b) is halo or hydroxy, or R^(1a) and R^(1b) are fluoro, Het is a ring of formula (iii) where R is hydrogen, and R^(u) and R^(v) are as defined in the Summary and R⁵ and R⁹ are as defined in the Summary can prepared from compounds of formula 7 as illustrated in Scheme 2 above by proceeding under the reaction conditions described in Scheme 1 above.

Compounds of formula 1 are either commercially available or they can be prepared by methods known in the art. For example, compound of formula 7 (2R,3R,4R,5R)-2-(6-amino-2-chloro-9H-purin-9-yl)-5-(hydroxymethyl)-3-methyltetrahydrofuran-3,4-diol is commercially available or can be prepared according to procedure reported by Caballero, Gerado et al., Helvetica Chimica Acta, 85(5), 1284-1294; 2002 and by Li, Nan-Sheng et al., J. Org. Chem., 74(5), 2227-2230, 2009). (2R,3R,4R,5R)-2-(6-Amino-2-chloro-9H-purin-9-yl)-3-ethynyl-5-(hydroxymethyl)tetrahydrofuran-3,4-diol can be prepared according to procedures reported by Nadler, Andre and Diedrerichsen, Ulf., European Journal of Organic Chemistry, 9, 1544-1549; 2008. (2R,3R,4R,5R)-2-(6-Amino-2-chloro-9H-purin-9-yl)-5-(hydroxymethyl)-3-vinyltetrahydrofuran-3,4-diol can be prepared according to procedures reported by Blatt, Lawrence M. et al., U.S. Pat. Appl. Publ. No. 20150366888, and by Carroll, Steven S. et al., PCT Int. Appl., publication no. WO 2004000858. (2R,3R,4R,5R)-2-(6-Amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydro-furan-3-carbonitrile can be prepared according to procedures reported by Ohtawa, Masaki et al., J. Med. Chem, 50(9), 2007-2009; 2007.

(2R,3R,4R,5R)-5-(6-Amino-2-chloro-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)-4-methyltetrahydrofuran-3-ol is commercially available. (2R,3R,5R)-5-(6-Amino-2-chloro-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4,4-difluorotetrahydrofuran-3-yl benzoate can be prepared by reacting ((2R,3R)-3-(benzoyloxy)-4,4-difluoro-5-iodotetrahydrofuran-2-yl)methyl benzoate and 2-chloroadenine under the Vorbrueggen conditions [(i) (CH₃Si)₂NH, (NH₄)₂SO₄ or Me₃SiN═CMeOSiMe₃, (ii) TMSOTf, refluxing] according to the procedure reported by Vorbrueggen, Helmut & Ruh-Polenz, Carmen, Organic Reactions, 55, 2000 and by Beigelman, Leonid, et al., US patent application, publication No. US 2013/0165400, followed by removal of the benzoyl groups in the resulting compound (2R,3R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-2-((benzoyloxy)methyl)-4,4-difluorotetrahydrofuran-3-yl benzoate as described below.

Compounds of Formula (I) where Het is other than purine can be prepared from commercially available starting materials as described in Scheme 1 above.

Compounds of Formula (I having the structure:

where R^(1a) is alkyl, alkynyl, vinyl, or cyano, Het is a ring of formula (iii) where R^(s) is hydrogen, and R^(u) and R^(v) are as defined in the Summary and R⁵ and R⁹ are as defined in the Summary can be synthesized by proceeding as illustrated and described in a representative example in Scheme 3 below.

Selective protection of the 4′,5′-diol in a compound of formula 12 with 1,3-dichloro-1,1,3,3-tetraisopropyldisiloxane, followed by acylation of 2′-OH in the resulting compound 13 with a suitable reagent such as 1,1′-thiocarbonyldiimidazole, 0-phenyl chloro thioformate, or methyl chloro oxoacetate provides a compound of formula 14. Compound 14 is converted to a compound of formula 15 where R^(1b) is hydrogen via deoxygenation of 2′-OH. The deoxygenation reaction is carried out by heating 14 in the presence of Bu₃SnH and AIBN at high temperature e.g., in refluxing toluene. Protection of amino group in compound 15, followed by removal of the silyl protecting group provides a compound of formula 16 which is then converted into a compound of Formula (I) as described in Scheme 1 above.

Compound of formula (I) where R^(1a) is ethynyl can be converted to the corresponding compound of formula (I) where R^(1a) is vinyl via reduction of the ethynyl group with Lindlar catalyst in the presence of hydrogen.

Compounds of formula 12 such as (2R,3R,4R,5R)-2-(6-amino-2-chloro-9H-purin-9-yl)-5-(hydroxymethyl)-3-methyltetrahydrofuran-3,4-diol is commercially available. (2R,3R,4R,5R)-2-(6-Amino-2-chloro-9H-purin-9-yl)-5-(hydroxymethyl)-3-vinyltetrahydrofuran-3,4-diol can be prepared according to procedures reported by Blatt, Lawrence M. et al., U.S. Pat. Appl. Publ., No. 20150366888; or by Carroll, Steven S. et al., PCT Int. Appl., publication No. 2004000858; and (2R,3R,4R,5R)-2-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxy-5-(hydroxyl-methyl)tetrahydro-furan-3-carbonitrile can be prepared according to procedures reported by Ohtawa, Masaki et al., J. Med. Chem, 50(9), 2007-2009; 2007.

Compounds of Formula (I) having the structure:

where R^(1a) is alkyl, alkynyl, vinyl, or cyano, R^(1b) is hydroxy or halo; or R^(1a) and R^(1b) are fluoro; Het is a ring of formula (iii) and R^(u), R^(v), R⁵ and R⁹ are as defined in the Summary can be synthesized by proceeding as illustrated and described in a representative example in Scheme 4 below.

Selective protection of the primary alcohol in compound 17 with a silyl protecting group such as TBDPSCl, followed by acylation and removal of the 3′-OH in resulting compound 18 as described in Scheme 3 above provides a compound of formula 20. Treatment of compound 20 with Boc₂O, followed by removal of the silyl group provides a compound of formula 21 which is then converted to a compound of Formula (I) as described in Scheme 1 above. Compound of formula (I) where R^(1a) is ethynyl can be converted to the corresponding compound of formula (I) where R^(1a) is vinyl via reduction of the ethynyl group with Lindlar catalyst in the presence of hydrogen.

Compounds of formula 17 are either commercially available or they can be prepared by methods known in the art. For example, (2R,3R,4R,5R)-2-(6-amino-2-chloro-9H-purin-9-yl)-5-(hydroxymethyl)-3-methyltetrahydrofuran-3,4-diol is commercially available. (2R,3R,4R,5R)-2-(6-Amino-2-chloro-9H-purin-9-yl)-5-(hydroxymethyl)-3-vinyltetrahydrofuran-3,4-diol can be prepared according to procedures reported by Blatt, Lawrence M. et. al in U.S. Pat. Appl. Publ. No. 20150366888 or by Carroll, Steven S. et al, in PCT Int. Appl., Publication No., WO2004000858; and (2R,3R,4R,5R)-2-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxy-5-(hydroxymethyl)tetrahydro-furan-3-carbonitrile can be prepared according to procedures reported by Ohtawa, Masaki et al., J. Med. Chem, 50(9), 2007-2009; 2007.

Compounds of Formula (I) having the structure:

where R^(1a) is hydrogen or alkyl, R^(1b) is hydroxy or halo; R² is azido or NH₂ and R^(2b) is hydrogen, R³ is hydrogen, Het is a ring of formula (iii), and R^(u), R^(v), R⁵ and R⁹ are as defined in the Summary can be synthesized by proceeding as illustrated and described in a representative example in Scheme 5 below.

Compound 23 is synthesized by first converting the 3′-OH group in compound 22 to a ketone with a suitable oxidizing agent such as Dess-Martine periodinane, Swern, Moffet, PDC, or SO₃.pyridine, followed by reduction of the keto group with a suitable reducing agent such as sodium borohydride. Compound 22 can be prepared according to the procedure reported by Koole, L. H. et al., Acta Chemica Scandinavica, 43, 665-669, 1989. The 3′-OH group in compound 23 is then converted to the corresponding azide compound 24 by first converting 3′-OH to a suitable leaving group such as mesylate, tosylate or triflate under reaction conditions well known in the art, followed by displacement of the leaving group with sodium azide.

After protecting the 6-amino group in compound 24 with suitable protecting group such as tert-butoxycarbonyl, the 4-methoxytrityl group is selectively removed with an acid such as TFA to provide compound 25. Compound 25 is then converted to compound of formula 27 as described in Scheme 1 above. Sequential removal of the TBDMS and Boc groups in compound 27 is achieved with treatment with TBAF, followed by treatment with TFA to give a compound of Formula (I) where R^(2a) is azido and R⁹ is alkyl group which can then be converted to corresponding compounds of Formula (I) where R⁹ is hydrogen as described in Scheme 1 above. Compounds of Formula (I) where R^(2a) is azido can also be converted to corresponding compounds of Formula (I) where R^(2a) is amino under suitable hydrogenation reaction conditions such as H₂, Pd/C, Pd(OH)₂/C or Lindlar catalyst.

Compounds of Formula (I) having the structure:

where R^(1a) is azido or NH₂; Het is a ring of formula (iii) and R^(u), R^(v), R⁵ and R⁹ are as defined in the Summary can be synthesized by proceeding as illustrated and described in a representative example in Scheme 6 below.

Compound 28 is first converted to compound 30 according to the procedure described in Scheme 5 above. Compound 28 can be prepared according to the procedure reported by Secrist, John A., III et al., Journal of Medicinal Chemistry, 31(2), 405-10; 1988. Compound 31 is obtained from compound 30 via a 4-step process. The bis-silyl protecting group on the 3′,5′-diol of compound 30 is first removed by treatment of TBAF and then the primary alcohol is protected as a TBDPS ether. Further protection of the 3′-OH and 6-NH₂ groups with Boc₂O, followed by removal of the TBDPS ether with TBAF affords compound 31.

Compound 31 is converted to a compound of Formula (I) as described in Schemes 1 and 5 above.

Compounds of Formula (I) having the structure:

where R^(1a) is azido or NH₂, Het is a ring of formula (iii) where R^(s) is hydrogen, and R^(u), R^(u), R⁵ and R⁹ are as defined in the Summary can be synthesized by proceeding as illustrated and described in a representative example in Scheme 7 below.

Compounds of Formula (I) as shown above, from compound 32 by first converting it into compound 35 as described in Scheme 7. Compound 35 is then converted to a compound of Formula (I) by following the reaction conditions described in Scheme 1. Compound 32 can be prepared according to the procedure reported by Koole, L. H. et al., Acta Chemica Scandinavica, 43, 665-669, 1989.

Proceeding as described in Scheme 7 above, but substituting compound 32 with compounds (2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)-oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl)-tetrahydrofuran-3-ol,

and (2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-2-4-methoxyphenyl)diphenylmethoxy)methyl)-tetrahydrofuran-3-ol,

compounds of Formula (I) where

where R⁵ is as defined in the Summary respectively, can be synthesized.

(2R,3R,4R,5R)-5-(6-Amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl)-tetrahydrofuran-3-ol and (2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-2-(((4-methoxyphenyl)diphenylmethoxy)methyl)-tetrahydrofuran-3-ol which can be prepared according to reported procedure described in Koole, L. H. et al, Acta Chemica Scandinavica, 43, 665-669, 1989.

Compounds of Formula (I) having the structure:

where R^(1a) and R^(2a), or R^(1a) and R^(2b), or R^(1b) and R^(2b), or R^(1a), R^(2a), R^(1b) and R^(2b) are fluoro, Het is a ring of formula (iii), and R^(u), R^(v), R⁵ and R⁹ are as defined in the Summary can be synthesized by proceeding as illustrated and described in a representative example in Scheme 8 below.

Treatment of compound 36 with 2-chloro-6H-purin-6-amine according to the procedure reported by Sari, Ozkan. et al., Tetrahedron Letters, 58(7), 642-644; 2017 provides compound 37. Protection of the amino group with tert-butoxycarbonyl, followed by removal of the benzyl group provide primary alcohol compound 39 which is converted to a compound of Formula (I) as described in Scheme 1 above.

Compounds of Formula (I) having the structure:

where R^(2a) is alkyl, alkynyl, or cyano, Het is a ring of formula (iii) where R^(s) is hydrogen, and R^(u) and R^(v) and R⁵ is as defined in the Summary can be synthesized by proceeding as illustrated and described in a representative example in Scheme 9 below.

Selective protection of the primary alcohol in a compound of formula 40 with a suitable protecting group such tributyl diphenylsilyl, followed by treatment of the resulting silyl compound with Boc₂O under reaction conditions described above provides a compound of formula 41. Deoxygenation of the 3′-hydroxy under conditions described in Scheme 4 above provides a compound of formula 42 which is then converted into a compound of Formula (I) as described in Scheme 1 above.

Compound of formula 40 can be prepared by methods well known in the art. (2R,3S,4R,5R)-5-(6-Amino-2-chloro-9H-purin-9-yl)-2-(hydroxymethyl)-3-methyl-tetrahydrofuran-3,4-diol can be prepared according to the procedure reported by Franchetti, Palmarisa, et al., J. Med. Chem., 48(15), 4983-4989, 2005). (2R,3S,4R,5R)-5-(6-Amino-2-chloro-9H-purin-9-yl)-3-ethynyl-2-(hydroxymethyl)-tetrahydrofuran-3,4-diol can be prepared according to the procedure reported by Hulpia, Fabian et al., Bioorganic & Medicinal Chemistry Letters, 26(8), 1970-1972; 2016.

(2R,3S,4R,5R)-5-(6-Amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxy-2-(hydroxymethyl)tetrahydrofuran-3-carbonitrile can be prepared from (2R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)-oxy)-2-(((tert-butyldimethylsilyl)oxy)-methyl)dihydrofuran-3(2H)-one and NaCN according to the similar procedure reported by Camarasa, Maria Jose et al., Journal of Medicinal Chemistry, 32(8), 1732-8; 1989 or from 2-chloro-2′-arabino-fluoro-2′-deoxyadenosine according to the procedure reported by Ohtawa, Masaki et al., J. Med. Chem., 50(9), 2007-2010, 2007.

A compound of Formula (I):

where R^(1a) is hydrogen and R^(1b) is hydroxyl or R^(1a) is fluoro and R^(1b) is hydrogen, R³ is alkyl or aralkyl, Het is a ring of formula (iii) where R^(s) is hydrogen, and R^(u), R^(v), R⁵ and R⁹ are as defined in the Summary can be synthesized as illustrated and described in Scheme 10 below.

Conversion of primary alcohol group in a compound of formula 43 with a suitable oxidizing reagent such as Dess-Martin periodinane, Swern, Moffet, or PDC provides an aldehyde compound of formula 44. Compound 44 is reacted with a lithium or Grignard reagent of formula R³Li or R³MgX where R³ is alkyl or aralkyl and X is halo under conditions well known in the art to provide a compound of formula 45. Compound 45 is then converted to a compound of Formula (I) as described in Scheme 1 above.

The foregoing compound numbers refer soley to the genus structures in the above general synthesis section and not to the compounds disclosed elsewhere in the application.

SYNTHETIC EXAMPLES Example 1 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

Step 1:

To a solution of 2-chloro-2′-arabino-fluoro-2′-deoxyadenosine (3.00 g, 9.9 mmol) in DMF (10 mL) at 0° C. was added imidazole (1.68 g, 24.7 mmol) and followed by TBDPSCl (3.00 mL, 11.7 mmol) dropwise. The reaction mixture was allowed to warm up to room temperature and stirred further for 8 h before it was quenched with H₂O (50 mL) and extracted with EtOAc (3×50 mL). The combined organic layer was washed further with H₂O (2×100 mL), brine, dried over Na₂SO₄ and concentrated. The residue was purified by silica gel column chromatography (0-20% EtOAc in hexanes) to provide (2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-4-fluorotetrahydrofuran-3-ol (4.10 g).

Step 2:

To a solution of (2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-4-fluorotetrahydrofuran-3-ol (4.10 g, 7.56 mmol) in DMF (10 mL) at room temperature was added Et₃N (3.48 mL, 25.0 mmol), 4-DMAP (150 mg, 1.2 mmol) and Boc₂O (5.20 g, 23.8 mmol). The reaction mixture was stirred for 16 h before it was diluted with EtOAc (200 mL) and H₂O (100 mL). The organic layer was separated, washed with H₂O (2×100 mL), brine, dried over Na₂SO₄ and concentrated. The residue was purified by silica gel column chromatography (0-20% EtOAc in hexanes) to provide N6,N6-bis-(tert-butoxycarbonyl)-5′-O-(tert-butyldiphenylsilyl)-2′-arabino-fluoro-2′-deoxy-3′-O-(tert-butoxycarbonyl)-2-chloro-adenosine (5.58 g).

Step 3:

N6,N6-Bis-(tert-butoxycarbonyl)-5′-O-(tert-butyldiphenylsilyl)-2′-arabino-fluoro-2′-deoxy-3′-O-(tert-butoxycarbonyl)-2-chloro-adenosine (5.58 g, 6.6 mmol) was dissolved in THF (10 mL) at 0° C. and followed by addition of a solution of TBAF (10 mL, 10 mmol, 1 M in THF) dropwise. The reaction mixture was stirred from 0° C. to room temperature over 2.5 h before it was evaporated to dryness. The residue was purified by silica gel column chromatography (0-20% EtOAc in hexanes) to provide N6,N6-bis-(tert-butoxycarbonyl)-5′-O-(tert-butyldiphenylsilyl)-2′-arabino-fluoro-2′-deoxy-3′-O-(tert-butoxycarbonyl)-2-chloro-adenosine-fluoro-2′-deoxy-3′-O-(tert-butoxycarbonyl)-2-chloro-adenosine (3.15 g).

Step 4:

To a solution of N6,N6-bis-(tert-butoxycarbonyl)-2′-fluoro-2′-deoxy-3′-O-(tert-butoxycarbonyl)-2-chloro-adenosine (800 mg, 1.32 mmol) in toluene (2 mL) was added diethyl 2-diazomalonate (321 mg, 1.72 mmol) and Rh₂(OAc)₄ (59 mg, 0.13 mmol) under argon atmosphere. The resulting mixture was stirred at 95° C. for 2 h before it was allowed to cool to room temperature. The organic volatile was removed under reduced pressure. The resulting crude was purified by silica gel column chromatography (0-20% EtOAc in hexanes) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate as a foam (770 mg).

Step 5:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-malonate (300 mg, 0.394 mmol) in CH₂Cl₂ (2 mL) at 0° C. was added TFA (3 mL). The resulting mixture was allowed to warm up to room temperature and stirred for 2 h before it was concentrated under reduced pressure to provide crude diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate.

Step 6:

To a solution of crude diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate (0.394 mmol) in THF (2 mL) and H₂O (2 mL) at room temperature was added LiOH monohydrate (200 mg). The resulting mixture was stirred overnight before it was cooled to 0° C. and acidified to pH ˜6 with 1N HCl(aq) solution and concentrated under reduced pressure. The crude residue was purified by preparative reversed-phase HPLC to provide 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid as a white solid.

¹H NMR (D₂O, 300 MHz) δ 8.47 (d, J=1.8 Hz, 1H), 6.46 (dd, J=4.4, 13.1 Hz, 1H), 5.27 (t, J=8.5 Hz, 1H), 4.60-4.72 (m, 2H), 4.15 (q, J=4.6 Hz, 1H), 3.86-4.03 (m, 2H); LC/MS [M+H]=406.

Example 2 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (170 mg, 0.223 mmol) in DMF (2 mL) at 25° C. was added Cs₂CO₃ (145 mg, 0.446 mmol). The reaction mixture was stirred for 30 min and followed by addition of BnBr (53 uL, 0.446 mmol). The reaction mixture was stirred for 3.5 h before it was diluted with H₂O (20 mL) and extracted with EtOAc (3×30 mL). The combined organic layer was washed further with H₂O (2×40 mL), brine, dried over Na₂SO₄ and concentrated. The resulting crude was purified by silica gel column chromatography (0-16% EtOAc in hexanes) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluoro-tetrahydrofuran-2-yl)methoxy)-2-benzylmalonate as a foam.

Step 2:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (200 mg, 0.235 mmol) in CH₂Cl₂ (2 mL) at 0° C. was added TFA (2 mL). The resulting mixture was stirred at room temperature for 2.5 h before it was concentrated under reduced pressure to provide diethyl diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate.

Step 3:

To a solution of crude 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate in THF (2 mL) and H₂O (2 mL) at 0° C. was added LiOH monohydrate (150 mg). The resulting mixture was stirred at room temperature overnight before it was cooled to 0° C. and acidified to pH ˜6 with 1N HCl(aq) solution and concentrated under reduced pressure. The crude residue was purified by preparative reversed-phase HPLC to provide 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid as a white solid.

¹H NMR (D₂O, 300 MHz) δ 8.27 (bs, 1H), 8.16 (s, 1H), 7.11 (bs, 5H), 6.35 (dd, J=4.3, 13.5 Hz, 1H), 5.17-5.36 (m, 1H), 4.53-4.56 (m, 1H), 4.18-4.28 (m, 1H), 3.70-3.85 (m, 2H), 3.24 (s, 2H); LC/MS [M+H]=496.

Example 3 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with methyl 4-(bromomethyl)benzoate provided 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.33 (d, J=2 Hz, 1H), 7.78-7.81 (d, J=8.3 Hz, 2H), 7.38-7.41 (d, J=8.3 Hz, 2H), 6.41-6.47 (dd, J=4.5, 13.6 Hz, 1H), 5.07-5.28 (dt, J=4.1, 52 Hz, 1H), 4.64-4.73 (dt, J=4.2, 18.0 Hz, 1H), 4.15-4.19 (q, J=4.6 Hz, 1H), 3.93-4.10 (m, 2H), 3.42-3.55 (m, 2H); LC/MS [M−H]=538.

Example 4 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((6-chloropyridin-3-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 5-(bromomethyl)-2-chloropyridine, the title compound was obtained as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.27 (s, 1H), 8.24 (d, J=2.5 Hz, 1H), 7.74 (dd, J=2.3, 8.4 Hz, 1H), 7.22 (d, J=8.3 Hz, 2H), 6.43 (dd, J=4.3, 13.7 Hz, 1H), 5.15 (dt, J=4.8, 51.7 Hz, 1H), 4.60-4.70 (m, 1H), 4.15-4.18 (m, 1H), 3.97-4.09 (m, 2H), 3.35-3.42 (m, 2H); LC/MS [M+H]=531.

Example 7 Synthesis of 2-(((2R,3R,4R,5R)-5-(6-amino-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

2-(((2R,3R 4R,5R)-5-(6-Amino-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid was prepared from diethyl 2-(((2R,3R,4R,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-malonate according to the procedure for Example 2. The title compound was isolated as a white solid.

LC/MS [M+H]=462.

Examples 8a, 8b, Sc and 8d Synthesis of ((R)-1-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-ethoxy-2-oxoethyl)phosphonic acid; ((S)-1-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-ethoxy-2-oxoethyl)phosphonic acid; ((R)-1-(((2R,3R,4S,5R)-5-(6-((tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-ethoxy-2-oxoethyl)phosphonic acid; and ((S)-1-(((2R,3R,4S,5R)-5-(6-((tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-ethoxy-2-oxoethyl)phosphonic acid

Step 1:

To a solution of N6,N6-bis-(tert-butoxycarbonyl)-2′-arabino-fluoro-2′-deoxy-3′-O-(tert-butoxycarbonyl)-2-chloro-adenosine (512 mg, 0.85 mmol) in toluene (5 mL) was added ethyl 2-diazo-2-(diethoxyphosphoryl)acetate (37 mg, 1.10 mmol) and Rh₂(OAc)₄ (37 mg, 0.08 mmol) under argon atmosphere. The resulting mixture was stirred at 95° C. for 3 h before it was allowed to cool to room temperature. The organic volatile was removed under reduced pressure. The resulting crude was purified by silica gel column chromatography (5-100% EtOAc in hexanes) to provide ethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(diethoxyphosphoryl)acetate (557 mg) as a mixture of diastereomers.

Step 2:

To a solution of ethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(diethoxyphosphoryl)acetate (345 mg, 0.42 mmol) in MeCN (6 mL) at −20° C. was added TMSBr (441 mL, 3.34 mmol) dropwise. The resulting mixture was stirred from −20 to 0° C. over 26 h before it was quenched with H₂O (4 mL). The resulting mixture was stirred at 0° C. for 20 h before it was quenched with NH₄OH (6 mL) and concentrated under reduced pressure. The crude residue was purified by preparative reversed-phase HPLC to provide the title compounds as white solids.

((R)-1-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-ethoxy-2-oxoethyl)phosphonic acid (diastereisomer 1)

¹H NMR (CD₃OD, 300 MHz) δ 8.54 (bs, 1H), 6.42 (dd, J=4.1, 10.1 Hz, 1H), 5.09-5.31 (m, 1H), 4.55-4.74 (m, 1H), 4.37-4.53 (m, 1H), 4.27 (q, J=7.1 Hz, 2H), 4.10-4.20 (m, 1H), 3.84-4.01 (m, 2H), 1.28 (t, J=6.9 Hz, 3H); LC/MS [M+H]=470.

((S)-1-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-ethoxy-2-oxoethyl)phosphonic acid (diastereisomer 2)

¹H NMR (CD₃OD, 300 MHz) 8.52 (bs, 1H), 6.42 (dd, J=4.6, 12.9 Hz, 1H), 5.21 (dt, J=4.6, 52.3 Hz, 1H), 4.72 (dt, J=4.9, 13.1 Hz, 1H), 4.45 (d, J=18.4 Hz, 1H), 4.27 (q, J=7.1 Hz, 2H), 4.10-4.20 (m, 1H), 3.85-4.01 (m, 2H), 1.29 (t, J=7.0 Hz, 3H); LC/MS [M+H]=470.

((R)-1-(((2R,3R,4S,5R)-5-(6-((tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-ethoxy-2-oxoethyl)phosphonic acid (NHBoc diastereisomer 1)

¹H NMR (CD₃OD, 300 MHz) δ 8.93 (bs, 1H), 8.46 (bs, 1H), 6.56 (m, 1H), 5.07-5.32 (m, 1H), 4.65-4.74 (m, 1H), 4.40 (d, J=23 Hz, 1H), 4.27 (q, J=6.9 Hz, 2H), 4.11-4.20 (m, 1H), 3.85-4.05 (m, 2H), 1.59 (bs, 9H), 1.26-1.35 (m, 3H); LC/MS [M+H]=571.

((S)-1-(((2R,3R,4S,5R)-5-(6-((tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-ethoxy-2-oxoethyl)phosphonic acid (NHBoc diastereisomer 1)

¹H NMR (CD₃OD, 300 MHz) δ 8.95 (bs, 1H), 8.46 (bs, 1H), 6.53 (m, 1H), 5.15-5.39 (m, 1H), 4.65-4.74 (m, 1H), 4.40 (m, 1H), 4.17-4.42 (m, 2H), 4.11-4.17 (m, 1H), 3.85-3.95 (m, 2H), 1.59 (bs, 9H), 1.26-1.35 (m, 3H); LC/MS [M+H]=571.

Example 10 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-cyanobenzyl)malonic acid

Proceeding as described in Example 2 above, but substituting benzyl bromide with 4-(bromomethyl)-benzonitrile the title compound was prepared.

¹H NMR (CD₃OD, 300 MHz) δ 8.27 (s, 1H), 7.48 (s, 4H), 6.43-6.46 (m, 1H), 5.09-5.26 (m, 1H), 4.63-4.69 (m, 1H), 3.97-4.17 (m, 3H), 3.42-3.49 (m, 2H); LC/MS [M+H]=521.

Example 11 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-cyanobenzyl)malonic acid

Proceeding as described in Example 2 above, but substituting benzyl bromide with 3-(bromomethyl)benzonitrile, the title compound was obtained as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.27 (s, 1H), 7.67 (bs, 1H), 7.57-7.60 (d, J=8 Hz, 1H), 7.46-7.49 (d, J=8 Hz, 1H), 7.30-7.35 (t, J=8 Hz, 1H), 6.43-6.49 (dd, J=4, 13 Hz, 1H), 5.08-5.28 (dt, J=4, 50 Hz, 1H), 4.62-4.71 (dt, J=4, 17 Hz, 1H), 4.17-4.21 (q, J=4 Hz, 1H), 4.00-4.12 (m, 2H), 3.40-3.52 (m, 2H); LC/MS [M+H]=521.

Example 12 Synthesis of diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(trifluoromethoxy)benzyl)malonate

Proceeding as described in Example 2 above, but substituting benzyl bromide with 1-(bromomethyl)-4-(trifluoromethoxy)benzene, the title compound was isolated as a white solid.

¹H NMR (CDCl₃, 300 MHz) δ 8.13 (bs, 1H), 7.25-7.28 (d, J=9 Hz, 2H), 7.09-7.12 (d, J=8 Hz, 1H), 6.43-6.50 (dd, J=4, 16 Hz, 1H), 6.10 (bs, NH₂), 5.05-5.24 (dt, J=3, 52 Hz, 1H), 4.68-4.77 (dt, J=5, 19 Hz, 1H), 4.22-4.28 (m, 4H), 3.98-4.10 (m, 3H), 3.41 (bs, 2H), 1.25-1.30 (dd, J=1, 7 Hz, 6H); LC/MS [M+H]=636.

Example 13 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(trifluoromethoxy)benzyl)malonic acid, (R)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydrofuran-2-yl)methoxy)-3-ethoxy-3-oxo-2-(4-(trifluoromethoxy)-benzyl)propanoic acid and (S)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydrofuran-2-yl)methoxy)-3-ethoxy-3-oxo-2-(4-(trifluoromethoxy)-benzyl)propanoic acid

2-(((2R,9R,4S,5R)-5-(6-Amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(trifluoromethoxy)benzyl)malonic acid was prepared from diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(trifluoromethoxy)benzyl)-malonate via base hydrolysis with aq. LiOH in THF according to the procedure described in Example 2 as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.38 (s, 1H), 7.37-7.34 (d, J=8 Hz, 2H), 7.07-7.09 (d, J=8 Hz, 2H), 6.39-6.46 (dd, J=4, 15 Hz, 1H), 5.05-5.25 (dt, J=4, 52 Hz, 1H), 4.63-4.80 (m, 1H), 4.15-4.20 (m, 1H), 3.91-3.99 (m, 2H), 3.37 (bs, 2H); LC/MS [M+H]=580.

A pair of diastereoisomer of mono-ethyl ester: (R)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-ethoxy-3-oxo-2-(4-(trifluoromethoxy)benzyl)propanoic acid and (S)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydrofuran-2-yl)methoxy)-3-ethoxy-3-oxo-2-(4-(trifluoromethoxy)benzyl)-propanoic acid was prepared from the partial hydrolysis of compound of Example 12. The title compounds were purified by reversed-phase HPLC and isolated as white solids.

Polar diastereomer 1: ¹H NMR (CD₃OD, 300 MHz) δ 8.30 (s, 1H), 7.36-7.38 (d, J=8 Hz, 2H), 7.05-7.08 (d, J=8 Hz, 2H), 6.40-6.46 (dd, J=4, 14 Hz, 1H), 5.08-5.27 (dt, J=4, 52 Hz, 1H), 4.63-4.78 (m, 1H), 4.15-4.26 (m, 3H), 3.99-4.06 (m, 2H), 3.42-3.44 (m, 2H), 1.22-1.27 (t, J=7 Hz, 3H); LC/MS [M+H]=608.

Less polar diastereomer 2: ¹H NMR (CD₃OD, 300 MHz) δ 8.37 (s, 1H), 7.35-7.38 (d, J=8 Hz, 2H), 7.05-7.08 (d, J=8 Hz, 2H), 6.42-6.48 (dd, J=4, 14 Hz, 1H), 5.10-5.30 (dt, J=4, 52 Hz, 1H), 4.64-4.71 (m, 1H), 4.15-4.25 (m, 3H), 4.02-4.04 (d, J=4 Hz, 2H), 3.44 (s, 2H), 1.22-1.25 (t, J=7 Hz, 3H); LC/MS [M+H]=608.

Example 14 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(hydroxymethyl)benzyl)malonic acid

Step 1:

To a solution of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-formylbenzyl)malonic acid (200 mg, 0.23 mmol), prepared from diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate and 4-(bromomethyl)benzaldehyde according to the procedure described for Example 2, in EtOH at 0° C. was added NaBH₄ (11 mg, 0.29 mmol). The resulting mixture was stirred for 15 min before it was quenched with 1N HCl(aq) solution. The organic volatile was removed under reduced pressure and the aq. layer was extracted with EtOAc. The organic layer was washed with brine, dried (Na₂SO₄) and concentrated. The crude residue was purified by silica gel column chromatography (0-20% EtOAc in hexanes) to provide the corresponding benzyl alcohol.

Step 2-3:

The benzyl alcohol from the last step was converted to 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(hydroxymethyl)benzyl)malonic acid according to the procedure described for Example 2. The title compound was isolated as white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.29 (s, 1H), 7.26-7.29 (d, J=8 Hz, 2H), 7.14-7.29 (d, J=8 Hz, 2H), 6.40-6.46 (dd, J=4, 13 Hz, 1H), 5.08-5.28 (dt, J=4, 52 Hz, 1H), 4.63-4.72 (dt, J=4, 18 Hz, 1H), 4.49 (s, 2H), 4.14-4.18 (q, J=4 Hz, 1H), 3.96-4.08 (m, 2H), 3.33-3.42 (m, 2H); LC/MS [M+H]=526.

Example 16 Synthesis of 2-(((2R,3R,4S,5R)-((1H-tetrazol-5-yl)methyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

Step 1:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(cyanomethyl) (103 mg, 0.129 mmol) from the synthesis of Example 15 in toluene (5 mL) was added azidotrimethylsilane (234 uL, 1.8 mmol) in portions over period of 3 days followed by bis(tributyltin)oxide (20 uL, 0.0386 mmol), the reaction mixture was heated at 75° C. for three days before it was concentrated under reduced pressure. The crude was purified by silica gel column chromatography (0-4% MeOH in DCM) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(1H-tetrazol-5-ylmethyl)malonate.

Step 2:

Diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(1H-tetrazol-5-ylmethyl)malonate was converted to the title compound as a white solid by proceeding as described for Example 2 above.

¹H NMR (CD₃OD, 300 MHz) δ 8.58 (s, 1H), 6.44-6.50 (dd, J=4, 14 Hz, 1H), 5.09-5.29 (dt, J=4, 52 Hz, 1H), 4.59-4.68 (dt, J=4, 17 Hz, 1H), 4.19-4.22 (q, J=5 Hz, 1H), 4.10-4.12 (m, 2H), 3.83 (s, 2H); LC/MS [M+H]=488.

Example 17 Synthesis of 2-(3-(1H-tetrazol-5-yl)benzyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-azido-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid; 2-(((2R,3R,4S,5R)-5-(6-amino-2-azido-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-cyanobenzyl)malonic acid; and 2-(((2R,3R,4S,5R)-5-(2-azido-6-((tert-butoxycarbonyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-cyanobenzyl)malonic acid

Step 1:

A mixture of diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluoro-tetrahydrofuran-2-yl)methoxy)-2-(3-cyanobenzyl)malonate (230 mg, 0.26 mmol) (prepared as described in Example 2 above, by reacting diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate with 3-(bromomethyl)benzonitrile), NaN₃ (26 mg, 0.39 mmol) and NH₄Cl (18 mg, 0.34 mmol) in DMF (1.5 mL) was heated at 100° C. for 5 h before it was allowed to cool to room temperature. The crude reaction mixture was diluted with EtOAc and water. The aqueous layer was further extracted (2×) with EtOAc. The combined organic layer was washed with brine, dried (MgSO₄) and concentrated. The crude was purified by reversed-phase HPLC to provide diethyl 2-(3-(1H-tetrazol-5-yl)benzyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)-amino)-2-azido-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluoro-tetrahydrofuran-2-yl)methoxy)malonate and diethyl 2-(((2R,3R,4S,5R)-5-(2-azido-6-((tert-butoxycarbonyl)amino)-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(3-cyanobenzyl)malonate.

Step 2:

Diethyl 2-(3-(JH-tetrazol-5-yl)benzyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-azido-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate and diethyl 2-(((2R,3R,4S,5R)-5-(2-azido-6-((tert-butoxycarbonyl)amino)-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(3-cyanobenzyl)malonate were then converted to the corresponding title compounds according to the procedure described for Example 2.

2-(((2R,3R,4S,5R)-5-(6-amino-2-azido-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-cyanobenzyl)malonic acid

¹H NMR (CD₃OD, 300 MHz) δ 8.66 (s, 1H), 7.66 (s, 1H), 7.56-7.59 (d, J=8 Hz, 1H), 7.46-7.48 (d, J=7 Hz, 1H), 7.33-7.35 (t, J=7 Hz, 1H), 6.52-6.57 (dd, J=5, 11 Hz, 1H), 5.17-5.38 (dt, J=5, 52 Hz, 1H), 4.64-4.75 (dt, J=4, 17 Hz, 1H), 4.21-4.25 (m, 1H), 3.97-4.16 (m, 2H), 3.41-3.54 (m, 2H); LC/MS [M+H]=528.

2-(((2R,3R,4S,5R)-5-(2-azido-6-((tert-butoxycarbonyl)amino)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-cyanobenzyl)malonic acid

¹H NMR (CD₃OD, 300 MHz) δ 8.72 (s, 1H), 7.67 (s, 1H), 7.56-7.59 (d, J=8 Hz, 1H), 7.46-7.49 (d, J=8 Hz, 1H), 7.31-7.36 (t, J=8 Hz, 1H), 6.57-6.62 (dd, J=4, 13 Hz, 1H), 5.16-5.36 (dt, J=4, 52 Hz, 1H), 4.63-4.87 (dt, J=4, 17 Hz, 1H), 4.22-4.25 (m, 1H), 4.00-4.14 (m, 2H), 3.41-3.53 (m, 2H), 1.60 (s, 9H); LC/MS [M+H]=628.

2-(3-(1H-tetrazol-5-yl)benzyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-azido-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

¹H NMR (CD₃OD, 300 MHz) δ 8.19 (bs, 1H), 7.99 (bs, 1H), 7.77-7.79 (d, J=8 Hz, 1H), 7.49-7.52 (d, J=8 Hz, 1H), 7.36-7.41 (t, J=8 Hz, 1H), 6.32-6.38 (dd, J=5, 13 Hz, 1H), 5.05-5.25 (dt, J=4, 52 Hz, 1H), 4.66-4.75 (dt, J=4, 18 Hz, 1H), 4.17-4.22 (m, 1H), 3.98-4.14 (m, 2H), 3.45-3.57 (m, 2H); LC/MS [M+H]=564.

Example 18 Synthesis of 2-(((2S,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3-azido-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid; and 2-(((2S,3S,4R,5R)-3-amino-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a stirred suspension of chromium trioxide (2.59 g, 25.9 mmol) in CH₂Cl₂ (60 mL) was added pyridine (4.19 mL, 51.8 mmol) dropwise and followed by immediately addition of acetic anhydride (2.45 mL, 25.9 mmol). This brown slurry was allowed to stir at room temperature for 10 minutes. To this mixture was added a solution of (2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl)-tetrahydrofuran-3-ol (synthesized from 2-chloroadenosin according to the procedure reported by Koole, L. H. et al., Acta Chemica Scandinavica, 43, 665-669, 1989) (5.95 g, 8.64 mmol) in CH₂Cl₂ (36 mL). The resulting mixture was stirred at room temperature for 18 h before it was passed through a short silica plug using EtOAc as eluent. The filtrate was washed with EDTA (2×100 mL) and brine (100 mL). The organic layer was separated and concentrated. The residue was purified by column chromatography on silica gel to give (2R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl)-dihydrofuran-3(2H)-one (4.19 g, 71% yield).

Step 2:

A solution of (2R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl)-dihydrofuran-3(2H)-one (4.19 g, 6.11 mmol) in a mixture of EtOH (42 mL) and water (2.1 mL) was stirred for 20 minute at −5° C. before the addition of sodium borohydride (324 mg, 8.55 mmol). The reaction was stirred for 5 h before it was carefully quenched with 1N aq. HCl until its pH reached 5. The reaction mixture was partitioned with EtOAc (100 mL). The organic layer was separated and washed with water (3×50 mL), brine, dried (MgSO₄) and concentrated. The crude residue was purified by column chromatography on silica gel to give (2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxy-phenyl)diphenylmethoxy)methyl)tetrahydrofuran-3-ol (2.19 g, 52% yield).

Step 3:

To a solution of (2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxy-phenyl)diphenylmethoxy)methyl)-tetrahydrofuran-3-ol (2.19 g, 3.18 mmol) in CH₂Cl₂ (33 mL) at 0° C. under argon atmosphere was added pyridine (2.3 mL, 28.6 mmol) and 4-dimethylaminopyridine (1.13 g, 9.22 mmol). The reaction mixture was stirred for 15 minutes before addition of Tf₂O (0.803 mL, 4.77 mmol). The reaction was stirred at 0° C. for 15 minutes before it was allowed to warm up to ambient temperature and stirred further for 4 hours. The reaction mixture was quenched by addition of water (22 mL). The organic layer was separated, dried (MgSO₄) and concentrated. The crude residue was purified by column chromatography on silica gel to give (2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl)tetrahydrofuran-3-yl trifluoromethanesulfonate (2.00 g, 76% yield).

Step 4:

To a solution of (2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl)-tetrahydrofuran-3-yl trifluoromethanesulfonate (1.65 g, 2.01 mmol) in DMF (33 mL) at 0° C. was carefully added sodium azide (1.31 g, 20.1 mmol). The reaction was allowed to warm to ambient temperature and stirred for 6 hours before it was partitioned with EtOAc (100 mL). The organic layer was separated, washed with water (2×22 mL), brine, dried over MgSO₄ and concentrated. To the crude oil in THF (5 mL) was added Boc₂O (1.70 g, 7.64 mmol) and 4-DMAP (24 mg, 0.20 mmol). The reaction was stirred under argon for 13 h before it was concentrated and the crude residue was purified by column chromatography on SiO₂ to give tert-butyl (tert-butoxycarbonyl)(9-((2R,3R,4R,5S)-4-azido-3-((tert-butyldimethylsilyl)oxy)-5-(((4-methoxyphenyl)diphenylmethoxy)methyl)-tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate (1.19 g, 65% yield for 2 steps).

Step 5:

To a solution of tert-butyl (tert-butoxycarbonyl)(9-((2R,3R 4R,5S)-4-azido-3-((tert-butyldimethylsilyl)oxy)-5-(((4-methoxyphenyl)diphenylmethoxy)methyl)-tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate (1.19 g, 1.30 mmol) was in DCM (30 mL) under argon atmosphere at room temperature was added trifluoroacetic acid (0.483 mL, 6.50 mmol) dropwise. The reaction was stirred for 7 h before it was concentrated. The crude residue was purified by column chromatography on SiO₂ to give tert-butyl (tert-butoxycarbonyl)(9-((2R,3R,4R,5S)-4-azido-3-((tert-butyldimethylsilyl)oxy)-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate (548 mg, 69% yield).

Step 6:

A mixture of tert-butyl (tert-butoxycarbonyl)(9-((2R,3R,4R,5S)-4-azido-3-((tert-butyldimethylsilyl)oxy)-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate (385 mg, 0.60 mmol) and diethyl 2-diazomalonate (145 mg, 0.78 mmol) were charged into a 25 mL round bottom flask and azeotroped twice with toluene. The resulting oil was dissolved in toluene (3.9 mL) and followed by addition of rhodium(II) acetate dimer (27 mg, 0.06) mmol under argon atmosphere. The reaction mixture was heated at 75° C. for 3 h before it was concentrated. The crude residue was purified by column chromatography on SiO₂ to give diethyl 2-(((2S,3R,4R,5R)-3-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-tetrahydrofuran-2-yl)methoxy)malonate (234 mg, 48% yield).

Step 7:

To a solution of diethyl 2-(((2S,3R,4R,5R)-3-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-tetrahydrofuran-2-yl)methoxy)malonate (234 mg, 0.29 mmol) in DMF (2.3 mL) at 0° C. was added oven dried cesium carbonate (191 mg, 0.59 mmol). The resulting suspension was stirred for 30 minutes and followed by dropwise addition of benzyl bromide (70 μL, 0.59 mmol). The reaction was allowed to warm to ambient temperature and stirred for 14 h before it was partitioned between EtOAc (25 mL) and water (22 mL). The organic layer was separated and washed water (22 mL), brine, dried over magnesium sulfate and concentrated. The crude residue was purified by column chromatography on SiO₂ to give diethyl 2-(((2S,3R,4R,5R)-3-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (100 mg, 38% yield).

Step 8:

A solution of diethyl 2-(((2S,3R,4R,5R)-3-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (100 mg, 0.11 mmol) in EtOAc (2.0 mL) was purged three times with argon gas and followed by addition of Lindlar catalyst (15 mg, 15% by wt) under a blanket of argon gas. The reaction mixture was then purged (thrice) and stirred under 1 atmosphere of H₂ for 18 h. The reaction mixture was filtered through celite and rinsed with DCM (10 mL). The filtrate was concentrated to provide diethyl 2-(((2S,3R,4R,5R)-3-amino-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)-amino)-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (92 mg, 94% yield) which was used in the next step without further purification.

Step 9:

To a solution of diethyl 2-(((2S,3R,4R,5R)-3-amino-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)-amino)-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (92 mg, 0.11 mmol) in a mixture of DCM (2.3 mL) and water (92 μL) and followed by dropwise addition of TFA (0.92 mL). The reaction was stirred for 16 hours before it was evaporated to dryness and azeotroped with acetonitrile twice. The resulting oil was taken up in THF (1.5 mL) and 4M NaOH (106 μL, 0.42 mmol) was added at room temperature. The reaction was stirred after 10 h before it was concentrated to dryness. The crude residue was purified by reversed-phase HPLC to give 2-(((2S,3S,4R,5R)-3-amino-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid.

¹H NMR (CD₃OD, 300 MHz) δ 8.35 (s, 1H), 7.10-7.77 (m, 5H), 6.45-6.52 (dd, J=4, 10 Hz, 1H), 5.18-5.39 (dt, J=4.5, 52 Hz, 1H), 4.65-4.73 (dt, J=5, 17 Hz, 1H), 4.21-4.24 (m, 1H), 3.95-4.18 (m, 2H), 3.45-3.59 (m, 2H); LC/MS [M+H]=493.

Step 10:

2-(((2S,3S,4R,5R)-5-(6-Amino-2-chloro-9H-purin-9-yl)-3-azido-4-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid was prepared from diethyl 2-(((2S,3R,4R,5R)-3-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-tetrahydrofuran-2-yl)methoxy)malonate under the similar deprotection conditions described in Step 9.

¹H NMR (CD₃OD, 300 MHz) δ 8.87 (bs, 1H), 6.03 (d, J=6.4 Hz, 1H), 5.17-5.21 (t, J=5.6 Hz, 1H), 4.61 (bs, 1H), 4.50 (bs, 1H), 4.20 (bs, 1H), 3.81-3.91 (q, J=0 Hz, 2H); LC/MS [M+H]=429.

Example 19 Synthesis of 2-(((2R,3S,4R,5R)-4-amino-5-(6-amino-2-chloro-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a cooled (−70° C.) solution of oxalyl chloride (4.18 mL, 48.78 mmol) in dry dichloromethane (100 mL) under argon atmosphere was added a solution of dry DMSO (7.1 mL, 99.44 mmol) in dichloromethane (18 mL) dropwise. After stirring for 30 minutes, a solution of (6aR,8R,9R,9aS)-8-(6-amino-2-chloro-9H-purin-9-yl)-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-9-ol (10.21 g, 18.76 mmol), synthesized from 2-chloroadenosine according to the procedure reported by Secrist, John A., III et al.; (Journal of Medicinal Chemistry, 31, 405-10, 1988); and by Chen, Robert H. K. (U.S. Pat. No. 5,208,327), in a mixture of dichloromethane and THF (46 mL, 1:1/v:v) was added dropwise over a period of 30 minutes and followed by triethylamine (16 mL, 114.44 mmol) via a syringe. The cooling bath was removed and the mixture was stirred at room temperature for 1.5 h before chloroform (190 mL) and water (650 mL) were added and the pH of the mixture was adjusted to neutral with 2N aq. HCl solution. The organic layer was separated and the aqueous phase was further extracted with chloroform (2×190 mL). The combined organic phase was dried (MgSO₄) and filtered through a short plug of celite. The filtrate was concentrated to provide (6aR,8R,9aR)-8-(6-amino-2-chloro-9H-purin-9-yl)-2,2,4,4-tetraisopropyldihydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-9(6aH)-one as a solid (8.37 g) which was used without further purification in the next step.

Step 2:

To a suspension of NaBH₄ (2.51 g, 66.4 mmol) in dry THF (130 mL) under argon atmosphere at 0° C. was added glacial acetic acid (19.3 mL, 337 mmol) dropwise. The reaction mixture was stirred for 1.5 h and followed by addition of a solution of (6aR,8R,9aR)-8-(6-amino-2-chloro-9H-purin-9-yl)-2,2,4,4-tetraisopropyldihydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-9(6aH)-one (3 g, 5.53 mmol) in tetrahydrofuran (30 mL) via a syringe. The mixture was stirred for 4 h at 0° C. The organic volatile was removed under reduced pressure and the residue was partitioned with EtOAc (115 mL) and saturated aqueous sodium bicarbonate (115 mL). The organic phase was separated and the aqueous layer was further extracted with EtOAc (2×100 mL). The combined organic phase was dried (MgSO₄), filtered and concentrated to provide (6aR,8R,9R,9aS)-8-(6-amino-2-chloro-9H-purin-9-yl)-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-9-ol as a foam (2.69 g).

Step 3:

To a solution of (6aR,8R,9R,9aS)-8-(6-amino-2-chloro-9H-purin-9-yl)-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-9-ol (2.297 g, 4.22 mmol) in anhydrous dichloromethane (90 mL) under argon atmosphere was added pyridine (3.1 mL, 38 mmol) and 4-dimethylaminopyridine (1.5 g, 12.24 mmol). The mixture was cooled to 0° C. and stirred for 15 minutes and followed by addition of trifluoromethanesulfonic anhydride (1.1 mL, 6.33 mmol) dropwise. After stirring for 5 minutes, the cooling bath was removed and the mixture was stirred at room temperature for 3 hours. The reaction was quenched with cold water (90 mL). The organic phase was separated and the aqueous phase was further extracted with DCM (2×80 mL). The combined organic phase was dried (MgSO₄), filtered and concentrated. The crude residue was purified by column chromatography on silica gel (25-85% EtOAc in hexanes) to provide (6aR,8R,9S,9aR)-8-(6-amino-2-chloro-9H-purin-9-yl)-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]-trioxadisilocin-9-yl trifluoromethanesulfonate as a white solid (1.9 g).

Step 4:

To a solution of (6aR,8R,9S,9aR)-8-(6-amino-2-chloro-9H-purin-9-yl)-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-9-yl trifluoromethanesulfonate (1.9 g, 2.8 mmol) in anhydrous DMF (12 mL) under argon atmosphere was added sodium azide (1.83 g, 28.1 mmol). The mixture was stirred for 24 hours before it was partitioned with EtOAc (120 mL) and brine (120 mL). The organic phase was separated and the aqueous phase was further extracted with EtOAc (2×100 mL). The combined organic phase was dried (MgSO₄), filtered and concentrated to provide 9-((6aR,8R,9R 9aS)-9-azido-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-8-yl)-2-chloro-9H-purin-6-amine as a solid (1.48 g).

Step 5:

To a solution of 9-((6aR,8R,9R,9aS)-9-azido-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-8-yl)-2-chloro-9H-purin-6-amine (1.48 g, 2.59 mmol) in anhydrous THF (31 mL) under argon atmosphere was added 4-dimethylaminopyridine (153 mg, 1.25 mmol) and then di-tert-butyl dicarbonate (1.2 g, 5.5 mmol). The mixture was stirred at room temperature for 16 hours before it was partitioned with EtOAc (40 mL) and brine (60 mL). The organic phase was separated and the aqueous phase was further extracted with EtOAc (2×40 mL). The combined organic phase was dried (MgSO₄), filtered and concentrated. The crude product was purified by column chromatography on silica gel (15% EtOAc in hexanes) to provide tert-butyl (tert-butoxycarbonyl)(9-((6aR,8R 9R,9aS)-9-azido-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-8-yl)-2-chloro-9H-purin-6-yl)di-carbamate as a viscous oil (1.857 g).

Step 6:

To a solution of tert-butyl (tert-butoxycarbonyl)(9-((6aR,8R,9R,9aS)-9-azido-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-8-yl)-2-chloro-9H-purin-6-yl)di-carbamate (1.85 g, 2.41 mmol) in anhydrous THF (13 mL) under argon atmosphere at 0° C. was added a solution of tetrabutylammonium fluoride (6.3 mL, 1.0 M in THF, 6.3 mmol) dropwise. The flask was stirred overnight at 6° C. before it was partitioned with EtOAc (50 mL) and brine (50 mL). The organic phase was separated and the aqueous phase was further extracted with EtOAc (2×40 mL). The combined organic phase was dried (MgSO₄), filtered and concentrated. The crude product was purified by column chromatography on silica gel (55% EtOAc in hexanes) to provide tert-butyl (tert-butoxycarbonyl)(9-((2R,3R,4S,5R)-3-azido-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate as a solid (1.117 g).

Step 7:

To a solution of tert-butyl (tert-butoxycarbonyl)(9-((2R,3R,4S,5R)-3-azido-4-hydroxy-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate (408 mg, 0.77 mmol) and Rh₂(OAc)₄ (14 mg, 0.04 mmol) in dry toluene (2 mL) under argon atmosphere at room temperature was added diethyl 2-diazomalonate (173 mg, 0.93 mmol). The resulting mixture was stirred at 75° C. for 3 hours before it was allowed to cool to ambient temperature. The reaction mixture was then partitioned with EtOAc (20 mL) and brine (20 mL). The organic phase was separated and the aqueous phase was further extracted with EtOAc (2×20 mL). The combined organic phases was dried (MgSO₄), filtered and concentrated. The residue was purified by preparative thin layer chromatographic plates eluting with 60% EtOAc in hexanes to provide diethyl 2-(((2R,3S,4R,5R)-4-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate as a viscous oil (167 mg).

Step 8:

To a solution of diethyl 2-(((2R,3S,4R,5R)-4-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate (167 mg, 0.24 mmol) in dry DMF (1.5 mL) under argon atmosphere was added cesium carbonate (119 mg, 0.37 mmol) and then benzyl bromide (44 mg, 0.26 mmol). Additional amount of Cs₂CO₃ (60 mg) and BnBr (40 mg) were added to the reaction over 1.5 h and stirred for addition 1 h. The reaction mixture was then partitioned with EtOAc (20 mL) and brine (20 mL). The organic phase was separated and the aqueous phase was further extracted with EtOAc (2×20 mL). The combined organic phase was dried (MgSO₄), filtered and concentrated. The residue was purified by preparative thin layer chromatography (eluting with 40% EtOAc in hexanes) to provide diethyl 2-(((2R,3S,4R,5R)-4-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate as a viscous oil.

Step 9:

To a solution of diethyl 2-(((2R,3S,4R,5R)-4-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (101 mg, 0.13 mmol) in ethanol (4 mL) was added Lindlar's catalyst (25 mg) and then stirred under 1 atmosphere of H₂ for 18 hours. The reaction mixture was filtered through a plug of celite and rinsed with methanol. The filtrate was concentrated to provide diethyl 2-(((2R,3S,4R,5R)-4-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate as the product (100 mg).

Step 10:

To a solution of diethyl 2-(((2R,3S,4R,5R)-4-azido-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (100 mg, 0.13 mmol) in THF (1.5 mL) and water (0.75 mL) was added lithium hydroxide mono-hydrate (65 mg, 1.56 mmol). The material was vigorously stirred for 5 hours at ambient temperature and then stirred for 66 hours at 6° C. before it was cooled to 0° C. and followed by added a solution of 1N aq. HCl (1.7 mL). The reaction mixture was concentrated to provide an off-white powder (66 mg). This material was then taken up in a solution of trifluoroacetic acid in dichloromethane (1.5 mL, 1:1/v:v) and stirred for 1 h before it was concentrated. The residue was taken up in dichloromethane (5×5 mL) and concentrated each time. The crude was purified by reversed-phase HPLC to provide 2-(((2R,3S,4R,5R)-4-amino-5-(6-amino-2-chloro-9H-purin-9-yl)-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid (18 mg) as a solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.29 (s, 1H), 7.26-7.16 (m, 5H), 6.36 (d, J=9 Hz, 1H), 4.6 (d, J=5 Hz, 1H), 4.53-4.47 (m, 1H), 4.43 (bs, 1H), 4.01 (dd, J=2, 11 Hz, 1H), 3.80 (dd, J=2, 10 Hz, 2H), 3.46 (bs, 2H); LC/MS [M+H]=493.

Example 21 Synthesis of 2-(((2R,3R,4S,5R)-5-(4-(benzylamino)-1H-benzo[d]imidazol-1-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-carboxybenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with methyl 3-(bromomethyl)benzoate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.31 (s, 1H), 7.99 (s, 1H), 7.81-7.84 (d, J=7 Hz, 1H), 7.52-7.55 (d, J=71 Hz, 1H), 7.25-7.30 (t, J=8 Hz, 1H), 6.40-6.46 (dd, J=4, 13 Hz, 1H), 5.07-5.27 (dt, J=4, 52 Hz, 1H), 4.66-4.73 (dt, J=5, 17 Hz, 1H), 4.16-4.20 (m, 1H), 3.99-4.11 (m, 2H), 3.40-3.55 (m, 2H); LC/MS [M+H]=540.

Example 22 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-fluorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 3-fluorobenzyl bromide, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.33 (s, 1H), 7.03-7.18 (m, 3H), 6.81-6.82 (m, 1H), 6.41-6.47 (dd, J=4, 13 Hz, 1H), 5.09-5.29 (dt, J=4, 52 Hz, 1H), 4.65-4.74 (dt, J=5, 18 Hz, 1H), 4.15-4.19 (q, J=4 Hz, 1H), 3.96-4.10 (m, 2H), 3.36-3.46 (m, 2H); LC/MS [M+H]=514.

Example 23 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-fluorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-fluorobenzyl bromide, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.33 (s, 1H), 7.26-7.30 (m, 2H), 6.81-6.87 (t, J=9 Hz, 2H), 6.41-6.47 (dd, J=4, 13 Hz, 1H), 5.09-5.29 (dt, J=4, 52 Hz, 1H), 4.65-4.74 (dt, J=5, 18 Hz, 1H), 4.14-4.20 (m, 1H), 3.95-4.09 (m, 2H), 3.33-3.45 (m, 2H); LC/MS [M+H]=514.

Example 24 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-methoxybenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 3-methoxybenzyl bromide, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.41 (s, 1H), 7.01-7.06 (t, J=7 Hz, 1H), 6.83-6.85 (m, 2H), 6.64-6.66 (d, J=8 Hz, 1H), 6.42-6.48 (dd, J=4, 11 Hz, 1H), 5.11-5.31 (dt, J=4, 52 Hz, 1H), 4.67-4.74 (dt, J=5, 18 Hz, 1H), 4.16-4.20 (m, 1H), 3.94-4.10 (m, 2H), 3.60 (d, J=1 Hz, 3H), 3.31-3.44 (m, 2H); LC/MS [M+H]=526.

Example 25 Synthesis of 2-(3-(1H-tetrazol-5-yl)benzyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

Step 1:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluoro-tetrahydrofuran-2-yl)methoxy)-2-(3-cyanobenzyl)malonate, prepared from diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate with 3-(bromomethyl)benzonitrile according to the procedure for Example 2, (380 mg, 0.43 mmol) in toluene (8 mL) was added azidotrimethylsilane (350 uL, 2.64 mmol) and bis(tributyltin)oxide (66 uL, 0.13 mmol) in portions over period of 4 days. The reaction mixture was then heated at 75° C. for four days before it was concentrated under reduced pressure. The crude residue was purified by column chromatography silica gel (0-4% MeOH in DCM) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluoro-tetrahydrofuran-2-yl)methoxy)-2-(3-1-H-tetrazol-5-ylbenzyl)malonate.

Step 2:

Diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluoro-tetrahydrofuran-2-yl)methoxy)-2-(3-1-H-tetrazol-5-ylbenzyl)malonate was then converted to the title compound as described in Example 2 above.

¹H NMR (CD₃OD, 300 MHz) δ 8.26 (s, 1H), 7.99 (s, 1H), 7.77-7.79 (d, J=7 Hz, 1H), 7.47-7.50 (d, J=8 Hz, 1H), 7.38 (t, J=7 Hz, 1H), 6.35-6.46 (m, 1H), 5.06-5.27 (m, 1H), 4.66-4.80 (m, 1H), 4.20-4.21 (m, 1H), 4.00-4.17 (m, 2H), 3.44-3.62 (m, 2H); LC/MS [M+H]=564.

Example 26 Synthesis of 2-(4-(1H-tetrazol-5-yl)benzyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

The title compound was prepared as described in Example 25 above by substituting 3-(bromomethyl)benzonitrile with 4-(bromomethyl)benzonitrile, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.27 (s, 1H), 7.79-7.82 (d, J=8 Hz, 2H), 7.50-7.52 (d, J=8 Hz, 2H), 6.42-6.47 (dd, J=4, 13 Hz, 1H), 5.09-5.29 (dt, J=4, 53 Hz, 1H), 4.65-4.74 (dt, J=4, 17 Hz, 1H), 4.18-4.21 (m, 1H), 3.98-4.14 (m, 2H), 3.46-3.58 (m, 2H); LC/MS [M+H]=564.

Example 27 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(benzyloxy)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 1-(benzyloxy)-4-(chloromethyl)benzene, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.33 (s, 1H), 7.27-7.35 (m, 5H), 7.17-7.20 (d, J=8 Hz, 2H), 6.72-6.75 (d, J=9 Hz, 2H), 6.40-6.46 (dd, J=5, 12 Hz, 1H), 5.10-5.29 (dt, J=4, 52 Hz, 1H), 4.89 (s, 2H), 4.66-4.75 (dt, J=5, 18 Hz, 1H), 4.15-4.17 (m, 1H), 3.93-4.08 (m, 2H), 3.30-3.41 (dd, J=5, 14 Hz, 2H); LC/MS [M+H]=602.

Example 28 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-hydroxybenzyl)malonic acid

To a solution of -(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(benzyloxy)benzyl)malonic acid (Example 27) (280 mg) in EtOAc (10 mL) was added Pd/C (50 mg, 10% wt.). The resulting mixture was stirred under 1 atmosphere of H₂ for 1.5 h before it was filtered off. The filtrate was concentrated to provide the title compound as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.33-8.34 (d, J=2 Hz, 1H), 7.09-7.12 (d, J=9 Hz, 2H), 6.58-6.61 (d, J=8 Hz, 2H), 6.40-6.46 (dd, J=4, 14 Hz, 1H), 5.05-5.27 (dt, J=4, 52 Hz, 1H), 4.63-4.71 (dt, J=4, 33 Hz, 1H), 4.14-4.18 (q, J=5 Hz, 1H), 3.96-4.00 (m, 2H), 3.46 (bs, 2H); LC/MS [M+H]=512.

Example 29 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-hydroxybenzyl)malonic acid

Proceeding as described in Examples 2 and 28 above, but substituting benzyl bromide with 1-(benzyloxy)-3-(chloromethyl)benzene, the title compound was isolated as white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.37 (s, 1H), 6.94-6.99 (t, J=8 Hz, 1H), 6.76-6.78 (m, 2H), 6.56-6.59 (d, J=8 Hz, 1H), 6.41-6.46 (dd, J=3, 12 Hz, 1H), 5.08-5.27 (dt, J=4, 52 Hz, 1H), 4.66-4.73 (m, 1H), 4.11-4.20 (m, 1H), 3.92-4.07 (m, 2H), 3.35 (m, 2H); LC/MS [M+H]=512.

Example 30 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxy-2-fluorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with methyl 4-(bromomethyl)-3-fluorobenzoate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.25 (s, 1H), 7.51-7.60 (m, 3H), 6.38-6.44 (dd, J=4, 14 Hz, 1H), 5.05-5.25 (dt, J=5, 52 Hz, 1H), 4.62-4.70 (m, 1H), 4.12-4.16 (m, 1H), 3.95-4.05 (m, 2H), 3.54 (bs, 2H); LC/MS [M+H]=558.

Example 31 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxy-3-fluorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with methyl 4-(bromomethyl)-2-fluorobenzoate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.32 (bs, 1H), 7.69-7.74 (t, J=8 Hz, 1H), 7.12-7.16 (m, 2H), 6.41-6.47 (dd, J=5, 14 Hz, 1H), 5.06-5.28 (dt, J=5, 52 Hz, 1H), 4.65-4.73 (m, 1H), 4.14-4.19 (m, 1H), 3.92-4.04 (m, 2H), 3.44 (bs, 2H); LC/MS [M+H]=558.

Example 32 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((5-(trifluoromethyl)furan-2-yl)methyl)-malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 2-(bromomethyl)-5-(trifluoromethyl)furan, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.38 (s, 1H), 6.78-6.79 (m, 4H), 6.39-6.45 (m, 2H), 5.06-5.26 (dt, J=4, 53 Hz, 1H), 4.62-4.71 (dt, J=4, 17 Hz, 1H), 4.13-4.16 (m, 1H), 3.97-4.00 (m, 2H), 3.58 (bs, 2H); LC/MS [M+H]=554.

Example 33 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((6-(trifluoromethyl)pyridin-3-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 5-(bromomethyl)-2-(trifluoromethyl)pyridine, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.60 (bs, 1H), 8.37 (s, 1H), 7.89 (d, J=9 Hz, 1H), 7.60 (d, J=8 Hz, 1H), 6.51-6.59 (dd, J=2, 22 Hz, 1H), 5.08-5.27 (dt, J=4, 53 Hz, 1H), 4.62-4.71 (dt, J=5, 17 Hz, 1H), 4.21-4.24 (m, 1H), 3.96-4.17 (m, 2H), 3.45-3.59 (q, J=15 Hz, 2H); LC/MS [M+H]=565.

Example 34 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-fluoro-4-(trifluoromethyl)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-(bromomethyl)-2-fluoro-1-(trifluoromethyl)benzene, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.76 (s, 1H), 7.27-7.49 (m, 3H), 6.51-6.56 (dd, J=4, 10 Hz, 1H), 5.18-5.39 (dt, J=5, 52 Hz, 1H), 4.62-4.71 (dt, J=5, 17 Hz, 1H), 4.21-4.24 (m, 1H), 3.96-4.17 (m, 2H), 3.45-3.59 (q, J=15 Hz, 2H); LC/MS [M+H]=582.

Example 35 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((3-phenylisoxazol-5-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 5-(bromomethyl)-3-phenylisoxazole, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.96 (s, 1H), 7.40-7.80 (m, 5H), 6.51-6.56 (dd, J=4, 10 Hz, 1H), 6.29 (s, 1H), 5.18-5.39 (dt, J=5, 52 Hz, 1H), 4.60-4.69 (dt, J=5, 17 Hz, 1H), 4.21-4.24 (m, 1H), 3.98-4.19 (m, 2H), 3.45-3.59 (q, J=15 Hz, 2H); LC/MS [M+H]=563.

Example 36 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((1-benzyl-1H-pyrazol-4-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 1-benzyl-4-(bromomethyl)-1H-pyrazole, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.45 (s, 1H), 7.56 (s, 1H), 7.414 (s, 1H), 7.17-7.23 (m, 3H), 7.00-7.03 (m, 2H), 6.39-6.45 (dd, J=4, 12 Hz, 1H), 5.10-5.31 (m, 3H), 4.65-4.75 (dt, J=5, 18 Hz, 1H), 4.14-4.17 (q, J=5 Hz, 1H), 3.93-4.02 (m, 2H), 3.30-3.36 (m, 2H); LC/MS [M+H]=576.

Example 37 Synthesis of 2-((1H-pyrazol-4-yl)methyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-1-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

2-((1H-pyrazol-4-yl)methyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid was prepared from Example 36 (200 mg) via de-benzylation with Pd/C (50 mg, 10 wt %) in an mixture of EtOH (0.5 mL) and EtOAc (5 mL) under 1 atmosphere of H₂. The catalyst was filtered off after 1 h and the filtrate was concentrated to provide the title compound as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.46 (s, 1H), 7.55 (s, 2H), 6.44-6.49 (dd, J=5, 12 Hz, 1H), 5.12-5.31 (dt, J=4, 53 Hz, 1H), 4.68-4.74 (dt, J=5, 13 Hz, 1H), 4.17-4.22 (q, J=5 Hz, 1H), 3.96-4.03 (m, 2H), 3.36 (bs, 2H); LC/MS [M+H]=486.

Example 38 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2-carboxythiazol-5-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with methyl 4-(bromomethyl)thiazole-2-carboxylate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.38 (d, J=2 Hz, 1H), 7.37-7.59 (d, J=4 Hz, 1H), 6.41-6.48 (m, 1H), 5.05-5.25 (dt, J=3, 52 Hz, 1H), 4.65-4.74 (m, 1H), 4.22-4.24 (m, 1H), 3.98 (bs, 2H), 3.66 (bs, 2H); LC/MS [M+H]=548.

Example 39 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2-carboxythiazol-4-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with methyl 5-(bromomethyl)thiazole-2-carboxylate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.48 (bs, 1H), 7.64 (bs, 1H), 6.41-6.44 (dd, J=5, 8 Hz, 1H), 5.14-5.33 (dt, J=4, 52 Hz, 1H), 4.73-4.79 (m, 1H), 4.01-4.04 (m, 1H), 3.70-3.79 (m, 2H), 3.73 (bs, 2H); LC/MS [M+H]=547.

Example 40 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((5-carboxy-1-methyl-1H-pyrazol-3-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with ethyl 5-(bromomethyl)-1-methyl-1H-pyrazole-3-carboxylate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.90 (bs, 1H), 6.51-6.56 (dd, J=4, 10 Hz, 1H), 6.35 (s, 1H), 5.19-5.40 (dt, J=4.7, 52 Hz, 1H), 4.61-4.70 (dt, J=5, 17 Hz, 1H), 4.21-4.24 (m, 1H), 3.96-4.17 (m, 2H), 3.93 (s, 3H), 3.45-3.59 (q, J=15 Hz, 2H); LC/MS [M+H]=543.

Example 41 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((3-carboxyisoxazol-5-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with ethyl 5-(bromomethyl)isoxazole-3-carboxylate, The title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.98 (bs, 1H), 6.51-6.56 (dd, J=4, 10 Hz, 1H), 6.39 (s, 1H), 5.18-5.39 (dt, J=5, 52 Hz, 1H), 4.62-4.71 (dt, J=5, 17 Hz, 1H), 4.21-4.25 (m, 1H), 3.97-4.17 (m, 2H), 3.43-3.56 (q, J=15 Hz, 2H); LC/MS [M+H]=531.

Example 42 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-cyano-3-fluorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-(bromomethyl)-2-fluorobenzonitrile, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.35 (bs, 1H), 7.04-7.57 (m, 3H), 6.48-6.54 (dd, J=4, 10 Hz, 1H), 5.18-5.39 (dt, J=5, 52 Hz, 1H), 4.65-4.75 (dt, J=5, 17 Hz, 1H), 4.21-4.24 (m, 1H), 3.94-4.18 (m, 2H), 3.45-3.60 (m, 2H); LC/MS [M+H]=539.

Example 43 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((5-carboxythiophen-3-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with ethyl 4-(bromomethyl)thiophene-2-carboxylate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.40 (bs, 1H), 7.73 (s, 1H), 7.50 (s, 1H), 6.44-6.49 (dd, J=3, 11 Hz, 1H), 5.03-5.30 (m, 1H), 4.65-4.75 (m, 1H), 4.15-4.23 (m, 1H), 3.96-4.10 (m, 2H), 3.39-3.53 (m, 2H); LC/MS [M+H]=546.

Example 44 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((5-carboxythiophen-2-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with ethyl 5-(bromomethyl)thiophene-2-carboxylate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.36 (s, 1H), 7.52-7.53 (d, J=4 Hz, 1H), 6.99-7.00 (d, J=4 Hz, 1H), 6.41-6.47 (dd, J=4, 14 Hz, 1H), 5.07-5.26 (dt, J=3, 48 Hz, 1H), 4.68-4.76 (dt, J=4, 18 Hz, 1H), 4.18-4.21 (m, 1H), 4.01-4.11 (m, 2H), 3.66 (s, 2H); LC/MS [M+H]=546.

Example 45 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(carboxymethyl)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with ethyl 2-(4-(bromomethyl)phenyl)acetate., the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.32 (bs, 1H), 7.24-7.26 (d, J=8 Hz, 2H), 7.08-7.10 (d, J=8 Hz, 2H), 6.40-6.46 (dd, J=4, 13 Hz, 1H), 5.07-5.27 (dt, J=4, 53 Hz, 1H), 4.63-4.72 (dt, J=4, 18 Hz, 1H), 4.15-4.20 (m, 1H), 3.95-4.06 (m, 2H), 3.44-3.54 (m, 2H), 3.40 (bs, 2H); LC/MS [M+H]=554.

Example 46 Synthesis of 2-([1,1′-biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid; (R)-2-([1,1′-biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-methoxy-3-oxopropanoic acid; (S)-2-([1,1′-biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-methoxy-3-oxopropanoic acid; (R)-2-([1,1′-biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-ethoxy-3-oxopropanoic acid; and (S)-2-([1,1′-biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-ethoxy-3-oxopropanoic acid

Step 1:

Diethyl 2-([1,1′-biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-N-(bis-(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (200 mg, 0.22 mmol), prepared by alkylating N6,N6-bis-(tert-butoxycarbonyl)-2′-arabino-fluoro-2′-deoxy-3′-O-(tert-butoxycarbonyl)-2-chloro-adenosine with 4-phenylbenzyl bromide according to the procedure described for Example 2, was dissolved in DCM (5 mL) and followed by addition of TFA (1.5 mL) at room temperature. The resulting mixture was stirred for 7 h before it was concentrated. The residue was re-taken up in DCM (3×5 mL) and concentrated under reduced pressure each time to remove the excessive TFA to provide diethyl 2-([1,1′-biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate.

Step 2:

To a solution of diethyl 2-([1,1′-biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (0.22 mmol) in a mixture of MeOH (2 mL), THF (2 mL) and H₂O (2 mL) was added LiOH.H₂O (90 mg, 2.15 mmol). The resulting mixture was stirred at room temperature for 5 h before the organic volatiles were removed under reduced pressure. The residue was re-dissolved in H₂O (2 mL) and acidified with 1N aq.HCl solution to pH 5 before it was concentrated under reduced pressure. The crude residue was purified by reversed-phase HPLC to provide the title compounds as white solids.

2-([1,1′-Biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

¹H NMR (CD₃OD, 300 MHz) δ 8.34 (bs, 1H), 7.27-7.46 (m, 9H), 6.41-6.46 (dd, J=5, 12 Hz, 1H), 5.09-5.30 (dt, J=4, 52 Hz, 1H), 4.68-4.77 (dt, J=4, 18 Hz, 1H), 4.15-4.19 (m, 1H), 3.97-4.12 (m, 2H), 3.43-3.52 (m, 2H); LC/MS [M+H]=572.

2-([1,1′-Biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-methoxy-3-oxopropanoic acid (Methyl diastereomer 1)

¹H NMR (CD₃OD, 300 MHz) δ 8.31 (bs, 1H), 7.26-7.48 (m, 9H), 6.42-6.47 (dd, J=5, 12 Hz, 1H), 5.10-5.31 (dt, J=4, 52 Hz, 1H), 4.67-4.77 (dt, J=5, 18 Hz, 1H), 4.15-4.19 (m, 1H), 3.98-4.10 (m, 2H), 3.78 (s, 3H), 3.40-3.52 (m, 2H); LC/MS [M+H]=586.

2-([1,1′-Biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-methoxy-3-oxopropanoic acid (Methyl diastereomer 2)

¹H NMR (CD₃OD, 300 MHz) δ 8.31 (bs, 1H), 7.25-7.47 (m, 9H), 6.40-6.46 (dd, J=4, 12 Hz, 1H), 5.10-5.30 (dt, J=4, 53 Hz, 1H), 4.66-4.76 (dt, J=5, 18 Hz, 1H), 4.08-4.17 (m, 2H), 3.92-3.96 (m, 1H), 3.79 (s, 3H), 3.40-3.52 (m, 2H); LC/MS [M+H]=586.

2-([1,1′-Biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-ethoxy-3-oxopropanoic acid (Ethyl diastereomer 1)

¹H NMR (CD₃OD, 300 MHz) δ 8.33 (bs, 1H), 7.26-7.48 (m, 9H), 6.44-6.49 (dd, J=5, 8 Hz, 1H), 5.11-5.31 (dt, J=4, 53 Hz, 1H), 4.71-4.77 (m, 1H), 4.17-4.25 (m, 3H), 4.00-4.09 (m, 2H), 3.44 (bs, 2H), 1.24-1.28 (t, J=7 Hz, 3H); LC/MS [M+H]=600.

2-([1,1′-Biphenyl]-4-ylmethyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-ethoxy-3-oxopropanoic acid (Ethyl diastereomer 2)

¹H NMR (CD₃OD, 300 MHz) δ 8.31 (bs, 1H), 7.25-7.47 (m, 9H), 6.40-6.46 (dd, J=5, 12 Hz, 1H), 5.10-5.30 (dt, J=4, 52 Hz, 1H), 4.68-4.77 (dt, J=5, 18 Hz, 1H), 4.22-4.29 (q, J=7 Hz, 2H), 4.10-4.19 (m, 2H), 3.93-3.98 (m, 1H), 3.40-3.53 (m, 2H), 1.25-1.30 (t, J=7 Hz, 3H); LC/MS [M+H]=600.

Example 47 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-chloro-4-methoxybenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-(bromomethyl)-2-chloro-1-methoxybenzene, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.34 (bs, 1H), 7.27-7.28 (d, J=2 Hz, 1H), 7.27-7.28 (dd, J=2, 8 Hz, 1H), 6.77-6.80 (d, J=8 Hz, 1H), 6.43-6.48 (dd, J=4, 11 Hz, 1H), 5.11-5.31 (dt, J=4, 52 Hz, 1H), 4.69-4.83 (dt, J=5, 17 Hz, 1H), 4.15-4.20 (m, 1H), 3.93-4.10 (m, 2H), 3.75 (s, 3H), 3.27-3.34 (m, 2H); LC/MS [M+H]=560.

Example 48 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-sulfamoylbenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-(bromomethyl)benzenesulfonamide, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.27 (s, 1H), 7.70-7.73 (d, J=8 Hz, 2H), 7.47-7.49 (d, J=8 Hz, 2H), 6.40-6.46 (dd, J=4, 14 Hz, 1H), 5.07-5.27 (dt, J=4, 52 Hz, 1H), 4.62-4.70 (dt, J=5, 17 Hz, 1H), 4.16-4.20 (q, J=5 Hz, 1H), 3.99-4.10 (m, 2H), 3.34-3.52 (m, 2H); LC/MS [M+H]=575.

Example 49 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-chloro-2-fluorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-chloro-2-fluorobenzyl bromide, the title compound was isolated as a white solid. [M+H]=548.

Example 50 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-((2-carboxyethyl)carbamoyl)-benzyl)malonic acid

Diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)((tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(4-((3-methoxy-3-oxopropyl)carbamoyl)-benzyl)malonate was prepared as described in Example 2 above but substituting benzyl bromide with methyl 3-(4-(bromomethyl)benzamido)propanoate. The title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.25 (s, 1H), 7.59-7.62 (d, J=8 Hz, 2H), 7.37-7.40 (d, J=8 Hz, 1H), 6.40-6.46 (dd, J=5, 14 Hz, 1H), 5.07-5.27 (dt, J=4, 52 Hz, 1H), 4.62-4.71 (dt, J=4, 18 Hz, 1H), 4.14-4.19 (m, 1H), 3.97-4.10 (m, 2H), 3.57-3.62 (t, J=7 Hz, 2H), 3.42-3.52 (m, 2H), 2.59-2.64 (t, J=7 Hz, 3H); LC/MS [M+H]=611.

Example 51 Synthesis of -(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2-carboxybenzofuran-5-yl)methyl)-malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with methyl 5-(bromomethyl)benzofuran-2-carboxylate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.21 (s, 1H), 7.64-7.67 (d, J=9 Hz, 2H), 7.59 (s, 1H), 7.39-7.42 (dd, J=1, 9 Hz, 1H), 7.14 (s, 1H), 6.45-6.49 (dd, J=7, 7 Hz, 1H), 5.16-5.38 (dt, J=6, 53 Hz, 1H), 4.75-4.85 (m, 1H), 4.18-4.25 (m, 2H), 3.88-3.93 (dd, J=3, 10 Hz, 2H), 3.45-3.64 (m, 2H); LC/MS [M+H]=580.

Example 52 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2,2,2-trifluoroethoxy)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 1-(bromomethyl)-4-(2,2,2-trifluoroethoxy)benzene, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.31 (s, 1H), 7.23-7.26 (d, J=9 Hz, 2H), 6.76-6.79 (d, J=8 Hz, 2H), 6.40-6.46 (dd, J=4, 13 Hz, 1H), 5.08-5.28 (dt, J=4, 52 Hz, 1H), 4.65-4.73 (dt, J=4, 17 Hz, 1H), 4.33-4.43 (m, 2H), 4.14-4.19 (m, 1H), 3.95-4.08 (m, 2H), 3.32-3.42 (m, 2H); LC/MS [M+H]=594.

Example 53 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(carboxymethoxy)benzyl)malonic acid

Step 1:

Diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(cyanomethoxy)benzyl)malonate was prepared according to the procedure described in Example 2 above but substituting benzyl bromide with 2-(4-(bromomethyl)phenoxy)acetonitrile.

Step 2:

Diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(cyanomethoxy)benzyl)malonate was then hydrolyzed to the corresponding acid with the treatment of aq. LiOH in THF. The title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.31 (s, 1H), 7.20-7.23 (d, J=8 Hz, 2H), 6.71-6.74 (d, J=8 Hz, 2H), 6.40-6.46 (dd, J=4, 13 Hz, 1H), 5.07-5.27 (dt, J=4, 52 Hz, 1H), 4.65-4.73 (m, 1H), 4.55 (s, 2H), 4.13-4.18 (m, 1H), 3.95-4.05 (m, 2H), 3.35-3.42 (m, 2H); LC/MS [M+H]=570.

Example 54 Synthesis of 2-benzyl-2-(((2R,3R,4S,5R)-5-(2-chloro-6-hydroxy-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

Step 1:

To a solution of ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(2,6-dichloro-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methyl benzoate (2.00 g, 3.77 mmol) in DMF was added K₂CO₃ (625 mg, 4.52 mmol) and 1-pentanol (655 μL, 6.03 mmol). The resulting mixture was stirred at 40° C. for 2 h before it was allowed to cool to room temperature and diluted with EtOAc and water. The organic layer was separated, washed with brine, dried (MgSO₄) and concentrated. This crude was dissolved in MeOH (20 mL) and cooled to 0° C. To this mixture was added a solution of NH₃ in MeOH (19 mL, 2.0 M in MeOH). The reaction mixture was stirred from 0° C. to ambient temperature over 18 h before it was concentrated under reduced pressure. The crude was purified by SiO₂ column chromatography (0-5% MeOH in DCM) to provide (2R,3R,4S,5R)-5-(2-chloro-6-(pentyloxy)-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-ol.

Step 2-4:

(2R,3R,4S,5R)-5-(2-chloro-6-(pentyloxy)-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-ol was then converted to diethyl 2-(((2R,3R,4S,5R)-3-((tert-butoxycarbonyl)oxy)-5-(2-chloro-6-(pentyloxy)-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate according to the procedure described for Example 1.

Step 5:

To a solution of diethyl 2-(((2R,3R,4S,5R)-3-((tert-butoxycarbonyl)oxy)-5-(2-chloro-6-(pentyloxy)-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (210 mg, 0.29 mmol) in DCM (4 mL) at room temperature was added TFA (2.0 mL). The reaction mixture was then stirred for 8 h before it was concentrated under reduced pressure. The residue was re-dissolved in DCM (2×5 mL) and re-concentrated. The crude product was dissolved in a mixture of THF (3 mL) and water (3 mL) and followed by addition of LiOH.H₂O (139 mg). The reaction was then stirred at room temperature for 48 h before it was concentrated under reduced pressure. The residue was re-dissolved in water (2 mL) and acidified to pH 6 with 1N aq. HCl solution before it was concentrated. The crude residue was purified by reversed-phase HPLC to provide the title compound as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.25 (s, 1H), 7.16-7.30 (m, 5H), 6.38-6.45 (dd, J=4, 15 Hz, 1H), 5.05-5.25 (dt, J=4, 52 Hz, 1H), 4.59-4.68 (dt, J=5, 17 Hz, 1H), 4.15-4.19 (q, J=5 Hz, 1H), 3.96-4.05 (m, 2H), 3.40 (bs, 2H); LC/MS [M+H]=497.

Example 55 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(5-methyl-1,3,4-oxadiazol-2-yl)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 2-(4-(bromomethyl)phenyl)-5-methyl-1,3,4-oxadiazole, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.26 (s, 1H), 7.66-7.69 (d, J=8 Hz, 2H), 7.37-7.39 (d, J=8 Hz, 2H), 6.42-6.47 (dd, J=5, 9 Hz, 1H), 5.14-5.33 (dt, J=4, 53 Hz, 1H), 4.62-4.70 (dt, J=5, 19 Hz, 1H), 4.15-4.20 (m, 2H), 3.92-3.96 (m, 1H), 3.32-3.52 (q, J=15 Hz, 2H), 2.61 (s, 3H); LC/MS [M+H]=578.

Example 56 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2′-cyano-[1,1′-biphenyl]-4-yl)-methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4′-(bromomethyl)-[1,1′-biphenyl]-2-carbonitrile, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.41 (d, J=1 Hz, 1H), 7.70-7.73 (dd, J=1, 8 Hz, 1H), 7.55-7.58 (dd, J=1, 8 Hz, 1H), 7.29-7.49 (m, 6H), 6.41-6.46 (dd, J=5, 11 Hz, 1H), 5.12-5.33 (dt, J=5, 53 Hz, 1H), 4.69-4.79 (dt, J=5, 18 Hz, 1H), 4.10-4.21 (m, 2H), 3.98-4.02 (m, 1H), 3.47-3.59 (m, 2H); LC/MS [M+H]=597.

Example 57 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((5-chlorobenzo[b]thiophen-3-yl)methyl)-malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 3-(bromomethyl)-5-chlorobenzo[b]thiophene, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.43 (s, 1H), 7.80 (s, 1H), 7.48-7.53 (m, 2H), 6.99-7.02 (d, J=8 Hz, 1H), 6.44-6.49 (dd, J=5, 10 Hz, 1H), 5.16-5.34 (m, 1H), 4.72-4.82 (dt, J=6, 18 Hz, 1H), 4.12-4.20 (m, 2H), 3.94-3.98 (m, 1H), 3.57-3.71 (m, 2H); LC/MS [M+H]=586.

Example 58 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(benzo[d]thiazol-2-ylmethyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 2-(bromomethyl)benzo[d]thiazole, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.35 (s, 1H), 7.80-7.86 (m, 2H), 7.28-7.41 (m, 2H), 6.37-6.42 (dd, J=4.5, 13 Hz, 1H), 5.08-5.11 (dt, J=4, 52 Hz, 1H), 4.72-4.81 (dt, J=4, 17 Hz, 1H), 4.08-4.19 (m, 3H), 3.99 (s, 2H); LC/MS [M+H]=553.

Example 59 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(methylcarbamoyl)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-(bromomethyl)-N-methylbenzamide, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.26 (s, 1H), 7.59-7.62 (d, J=8 Hz, 2H), 7.37-7.40 (d, J=8 Hz, 2H), 6.40-6.46 (dd, J=4, 13 Hz, 1H), 5.07-5.27 (dt, J=4, 52 Hz, 1H), 4.62-4.71 (dt, J=5, 18 Hz, 1H), 4.17-4.18 (q, J=4 Hz, 3H), 3.97-4.10 (m, 2H), 3.42-3.52 (m, 2H); LC/MS [M+H]=553.

Example 60 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl)benzyl)malonic acid

Step 1:

Dibenzyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate was prepared by proceeding as described in Example 1 above by, utilizing dibenzyl diazomalonate instead of diethyl diazomalonate.

Step 2:

Proceeding as described in Example 2 above, but utilizing methyl 4-(bromomethyl)benzoate in place of benzyl bromide and subsequent removal of Boc groups with treatment of TFA provided dibenzyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl)benzyl)-malonate.

Steps 3-4:

The title compound was isolated as a white solid from dibenzyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxy-tetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl)benzyl)malonate according to the procedure described for Example 28.

¹H NMR (CD₃OD, 300 MHz) δ 8.27 (s, 1H), 7.69-7.71 (d, J=8 Hz, 2H), 7.36-7.38 (d, J=8 Hz, 2H), 6.42-6.47 (dd, J=5, 11 Hz, 1H), 5.11-5.31 (dt, J=5, 53 Hz, 1H), 4.68-4.77 (dt, J=5, 18 Hz, 1H), 3.94-4.18 (m, 3H), 3.42-3.55 (m, 2H), 3.42-3.52 (m, 2H); LC/MS [M+H]=554.

Example 61 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-((E)-2-carboxyvinyl)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with methyl 3-(4-bromomethyl)cinnamate, the title compound was isolated as a white solid.

LC/MS [M+H]=566.

Example 62 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-carboxyethyl)benzyl)malonic acid

The title compound was prepared from 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-((E)-2-carboxyvinyl)benzyl)malonic acid (Example 58) by reducing the olefin bond with Pd/C in EtOH under 1 atmosphere of H₂. The title compound isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.38 (s, 1H), 8.30 (bs, 1H), 7.00-7.24 (m, 5H), 6.38-6.41 (dd, J=4, 10 Hz, 1H), 5.07-5.26 (dt, J=4.7, 52 Hz, 1H), 3.98-4.16 (m, 4H), 3.98-4.20 (m, 2H), 3.32-3.36 (m, 4H), 2.52-2.85 (m, 5H); LC/MS [M+H]=534.2.

Example 63 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-phenoxybenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 3-(phenoxy)benzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.38 (bs, 1H), 7.02-7.22 (m, 6H), 6.70-6.81 (m, 3H), 6.37-6.41 (dd, J=4.3, 13 Hz, 1H), 5.07-5.25 (m, 1H), 4.62-4.67 (m, 1H), 3.93-4.18 (m, 3H), 3.24 (m, 2H); LC/MS [M+H]=589.

Example 64 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-(trifluoromethoxy)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 3-trifluoromethoxybenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.34 (bs, 1H), 7.20-7.29 (m, 3H), 7.01-7.04 (d, 1H), 6.42-6.48 (dd, J=4, 13 Hz, 1H), 5.08-5.28 (dt, J=4.7, 52 Hz, 1H), 4.62-4.71 (dt, J=5, 17 Hz, 1H), 4.15-4.19 (m, 1H), 3.98-4.10 (m, 2H), 3.44-3.45 (m, 2H); LC/MS [M+H]=580.8.

Example 65 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-(trifluoromethyl)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 3-trifluoromethylbenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.96 (bs, 1H), 7.37-7.59 (m, 4H), 6.51-6.56 (dd, J=4, 10 Hz, 1H), 5.18-5.39 (dt, J=4.7, 52 Hz, 1H), 4.62-4.71 (dt, J=5, 17 Hz, 1H), 4.21-4.24 (m, 1H), 3.96-4.17 (m, 2H), 3.45-3.59 (q, J=15 Hz, 2H); LC/MS [M+H]=564.8.

Example 66 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(naphthalen-2-ylmethyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with naphthalene-2-ylmethyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.29 (bs, 1H), 7.60-7.72 (m, 3H), 7.51-7.53 (d, J=8 Hz, 1H), 7.36-7.39 (d, J=8 Hz, 1H), 7.19-7.30 (m, 2H), 6.40-6.45 (dd, J=4.7, 10.6 Hz, 1H), 5.10-5.30 (dt, J=4.3, 53 Hz, 1H), 4.71-4.77 (m, 1H), 4.09-4.20 (m, 1H), 3.91-4.12 (m, 2H), 3.49-3.62 (q, J=11 Hz, 2H); LC/MS [M+H]=546.9.

Example 67 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(trifluoromethyl)benzylmalonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-trifluoromethylbenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.31 (d, J=1.6 Hz, 1H), 7.41-7.49 (m, 4H), 6.40-6.46 (dd, J=4.3, 13.4 Hz, 1H), 5.08-5.29 (dt, J=4.0, 52 Hz, 1H), 4.63-4.88 (dt, J=3.9, 17.5 Hz, 1H), 4.15-4.19 (m, 1H), 3.99-4.10 (m, 2H), 3.49 (m, 2H); LC/MS [M+H]=564.8.

Example 68 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(methylsulfonyl)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-methylsulfonylbenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.28 (d, J=1 Hz, 1H), 7.55-7.75 (dd, J=8.3, 50 Hz, 2H), 6.40-6.46 (dd, J=4.6, 13.3 Hz, 1H), 5.08-5.26 (dt, J=4.1, 52 Hz, 1H), 4.63-4.68 (dt, J=4.9, 17.6 Hz, 1H), 4.15-4.19 (m, 1H), 3.99-4.10 (m, 2H), 3.52-3.53 (m, 2H), 3.03 (s, 3H); LC/MS [M+H]=574.9.

Example 69 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-(dimethylcarbamoyl)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 3-(bromomethyl)-N,N-dimethylbenzamide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.42 (bs, 1H), 7.20-7.41 (m, 4H), 6.41-6.47 (dd, J=4.4, 13.3 Hz, 1H), 5.08-5.28 (dt, J=4, 52 Hz, 1H), 4.59-4.68 (dt, J=4.3, 17.6 Hz, 1H), 4.14-4.19 (q, J=5 Hz, 1H), 4.02-4.05 (m, 2H), 3.52-3.53 (m, 2H), 3.00 (s, 3H), 2.92 (s, 3H); LC/MS [M+H]=568.

Example 70 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(thiophen-3-ylmethyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 3-(bromomethyl)thiophene, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.38 (bs, 1H), 7.19-7.22 (m, 2H), 7.02-7.04 (m, 1H), 6.40-6.46 (dd, J=4.3, 13.4 Hz, 1H), 5.07-5.27 (dt, J=3.9, 9, 52 Hz, 1H), 4.60-4.66 (m, 1H), 4.15-4.19 (q, J=5 Hz, 1H), 3.94-3.99 (m, 2H), 3.45 (bs, 2H); LC/MS [M+H]=503.

Example 71 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-aminobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with tert-butyl (3-(bromomethyl)phenyl)carbamate, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.33 (bs, 1H), 7.09-7.25 (bs, 4H), 6.39-6.44 (d, J=15 Hz, 1H), 5.06-5.24 (m, 1H), 4.18-4.28 (d, J=18.4 Hz, 1H), 4.20 (bs, 1H), 3.90 (bs, 2H), 3.39 (s, 2H); LC/MS [M+H]=512.

Example 72 Synthesis of 2-benzyl-2-(((2R,3R,4S,5R)-5-(6-(benzylamino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

Step 1:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(benzyl(tert-butoxycarbonyl)-amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluoro-tetrahydrofuran-2-yl)methoxy)malonate (130 mg, 0.173 mmol) in DMF (2 mL) at 25° C. was added Cs₂CO₃ (113 mg, 0.346 mmol). The reaction mixture was stirred for 30 min and followed by addition of benzyl bromide (41 uL, 0.346 mmol). The reaction mixture was stirred for 3.5 h before it was diluted with H₂O (15 mL) and extracted with EtOAc. The combined organic layer was washed further with H₂O, brine, dried over Na₂SO₄ and concentrated. The resulting crude was purified by silica gel column chromatography (0-20% EtOAc in hexanes) to provide diethyl 2-benzyl-2-(((2R,3R,4S,5R)-5-(6-(benzyl(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate as a foam.

Diethyl 2-benzyl-2-(((2R,3R,4S,5R)-5-(6-(benzyl(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate was converted into the title compound according to the procedure described for Example 2.

¹H NMR (CD₃OD, 300 MHz) δ 8.23 (bs, 1H), 7.23-7.42 (m, 10H), 6.36-6.45 (dd, J=2.6, 22.6 Hz, 1H), 4.96-5.14 (dd, J=2.4, 50 Hz, 1H), 4.74-4.80 (m, 3H), 4.16-4.19 (m, 1H), 3.73-3.88 (m, 2H), 3.35-3.40 (m, 2H); LC/MS [M+H]=587.

Example 73 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-hydroxybutyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with (4-bromobutoxy(tert-butyl)dimethylsilane, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.50-8.54 (m, 1H), 6.41-6.47 (dd, J=4.6, 12.7 Hz, 1H), 5.17-5.36 (dt, J=4.3, 52 Hz, 1H), 4.18-4.28 (dt, J=4.2, 17.3 Hz, 1H), 4.08-4.17 (m, 2H), 3.70-3.90 (m, 2H), 3.52 (t, J=6.3 Hz, 1H), 2.12-2.18 (m, 2H), 1.35-1.72 (m, 4H); LC/MS [M+H]=479.

Example 74 Synthesis of 1-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)ethane-1,1,2-tricarboxylic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with (ethyl 2-bromoacetate, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.78 (bs, 1H), 6.44-6.49 (dd, J=4.6, 11.8 Hz, 1H), 5.12-5.32 (dt, J=4.5, 52 Hz, 1H), 4.67-4.76 (m, 1H), 4.13-4.18 (q, J=4.7 Hz, 1H), 3.93-3.99 (m, 2H), 3.23 (s, 2H); LC/MS [M+H]=464.

Example 75 Synthesis of 2-(((2R,3R,4S,5R)-5-(4-(benzylamino)-1H-benzo[d]imidazol-1-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

Step 1:

A suspension of N6,N6-bis-Boc-5′-O-tert-butyldiphenylsilyl-3′-O-Boc-2-chloro-adenosine (1.00 g, 1.19 mol) in a mixture of t-BuOH and H₂O (8 mL, 1:1/v:v) was refluxed for 13 h before it was allowed to cool to room temperature and concentrated. The crude residue was purified by silica gel column chromatography (5-20% EtOAc in hexanes) to provide tert-butyl (9-((2R,3S,4R,5R)-4-((tert-butoxycarbonyl)oxy)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-3-fluorotetrahydro-furan-2-yl)-2-chloro-9H-purin-6-yl)carbamate.

Step 2:

To a solution tert-butyl (9-((2R,3S,4R,5R)-4-((tert-butoxycarbonyl)oxy)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-3-fluorotetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate (500 mg, 0.67 mmol) in MCCN (4 mL) at room temperature was added DBU (201 uL, 1.35 mmol) and followed by BnBr (119 uL, 0.996 mmol). The reaction mixture was stirred for 3 h before it was diluted with EtOAc (10 mL) and H₂O (10 mL). The organic layer was separated, washed with brine, dried over Na₂SO₄ and concentrated. The crude residue was purified by silica gel column chromatography (0-15% EtOAc in hexanes) to provide tert-butyl benzyl (9-((2R,3S,4R,5R)-4-((tert-butoxycarbonyl)oxy)-5-(((tert-butyldiphenylsilyl)oxy)-methyl)-3-fluorotetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate.

Step 3:

To a solution of tert-butyl benzyl (9-((2R,3S,4R,5R)-4-((tert-butoxycarbonyl)-oxy)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-3-fluorotetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate (545 mg, 0.655 mmol) was dissolved in THF (1 mL) at 0° C. and followed by addition of a solution of TBAF (1 mL, 0.982 mmol, 1 M in THF) dropwise. The reaction mixture was stirred from 0° C. to room temperature over 2.5 h before it was evaporated to dryness. The residue was purified by silica gel column chromatography (0-20% EtOAc in hexanes) to provide tert-butyl benzyl (9-((2R,3S,4R,5R)-4-((tert-butoxycarbonyl)oxy)-3-fluoro-5-(hydroxymethyl)tetrahydro-furan-2-yl)-2-chloro-9H-purin-6-yl)carbamate.

Step 4:

To a solution of tert-butyl benzyl (9-((2R,3S,4R,5R)-4-((tert-butoxycarbonyl)-oxy)-3-fluoro-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate (300 mg, 0.505 mmol) in toluene (3 mL) was added diethyl 2-diazomalonate (122 mg, 0.657 mmol) and Rh₂(OAc)₄ (22 mg, 0.051 mmol) under argon atmosphere. The resulting mixture was stirred at 80° C. for 2.5 h before it was allowed to cool to room temperature. The organic volatile was removed under reduced pressure. The resulting crude was purified by silica gel column chromatography (0-25% EtOAc in hexanes) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-(benzyl(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate.

Step 5:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(benzyl(tert-butoxycarbonyl)-amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydro-furan-2-yl)methoxy)malonate (200 mg, 0.27 mmol) in CH₂Cl₂ (2 mL) at 0° C. was added TFA (3 mL). The resulting mixture was warm up to room temperature and stirred for 2 h before it was concentrated under reduced pressure to provide crude diethyl 2-(((2R,3R,4S,5R)-5-(6-(benzylamino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate as a TFA salt which was used in the next step without further purification.

Step 6:

To a solution of crude diethyl 2-(((2R,3R,4S,5R)-5-(6-(benzylamino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate TFA salt (0.27 mmol) in THF (3 mL) and H₂O (1 mL) at room temperature was added LiOH monohydrate (50 mg). The resulting mixture was stirred overnight before it was cooled to 0° C. and acidified to pH ˜6 with 1N aq. HCl solution and concentrated under reduced pressure. The crude residue was purified by preparative reverse-phase HPLC to provide 2-(((2R,3R,4S,5R)-5-(4-(benzylamino)-1H-benzo[d]imidazol-1-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.40 (bs, 1H), 7.26-7.42 (m, 5H), 6.40-6.47 (dd, J=4.5, 13.8 Hz, 1H), 5.07-5.28 (dt, J=3.9, 52 Hz, 1H), 4.77 (bs, 2H), 4.62-4.70 (m, 2H), 4.13-4.16 (q, J=5 Hz, 1H), 3.90-3.98 (m, 2H); LC/MS [M+H]=497.

Example 76 Synthesis of ((R)-1-(((2R 3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(methylamino)-2-oxoethyl)phosphonic acid; and ((S)-1-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(methylamino)-2-oxoethyl)phosphonic acid

Step 1:

To a solution of diastereomeric mixture (1:1 ratio) of methyl 2-(((2R,3R,4S,5R)-5-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)-oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(dimethoxyphosphoryl)acetate (0.352 mmol) in THF (4 mL) at room temperature was added a solution of 1N aq. LiOH (0.7 mL). The resulting mixture was stirred for 75 minutes before it was cooled to 0° C. and acidified to pH ˜6 with 1N aq. HCl solution and concentrated under reduced pressure to provide a crude diastereomeric mixture of 2-(((2R,3R,4S,5R)-5-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluoro-tetrahydrofuran-2-yl)methoxy)-2-(dimethoxyphosphoryl)acetic acid (280 mg) which was used in the next step without further purification.

Step 2:

To a solution of the above crude diastereomeric mixture of 2-(((2R,3R,4S,5R)-5-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(dimethoxyphosphoryl)acetic acid (140 mg, 0.182 mmol) in dry DCM (10 mL) was added methylamine hydrochloride (49 mg, 0.73 mmol) and HATU (140 mg, 0.364 mmol). To this mixture was added DIEA (0.22 mL, 1.3 mmol) and the reaction mixture was stirred overnight. Additional amounts of methylamine (25 mg), HATU (140 mg) and DIEA (0.22 mL) were added and the mixture was stirred for 20 hours before it was quenched with water (25 mL). The organic layer was separated and the aqueous was extracted with DCM (3×25 mL). The combined organic phases were dried and the crude product was purified via preparative TLC (60% ethyl acetate/hexane) to provide methyl 2-(((2R,3R,4S,5R)-5-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(diethoxyphosphoryl)acetamide (66 mg) as a mixture of diastereomers (1:1 ratio).

Step 3:

Diastereomeric mixture of methyl 2-(((2R,3R,4S,5R)-5-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(diethoxyphosphoryl)acetamide was converted to a diastereomeric mixture of ((R)-1-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(methylamino)-2-oxoethyl)phosphonic acid and ((S)-1-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(methylamino)-2-oxoethyl)phosphonic acid as a solid (1:1 ratio) according to the procedure for Examples 8a and 8b.

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (1:1 ratio): S 8.63-8.91 (m, 1H), 6.51 (d, 1H, diastereomeric), 5.11-5.40 (m, 1H), 4.18-4.70 (m, 3H), 4.01 (broad s, 2H), 2.78 (broad d, 3H); LC/MS [M+H]=456.

Example 77 Synthesis of 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid

2-(((2R,3S,4R,5R)-5-(6-Amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid was prepared according to the procedure described for Examples 1 and 2. Isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.42 (bs, 1H), 7.79-7.82 (d, J=8.2 Hz, 2H), 7.34-7.37 (d, J=8.2 Hz, 2H), 6.02-6.04 (d, J=6.2 Hz, 1H), 4.74-4.78 (m, 1H), 4.30-4.35 (m, 2H), 3.80-4.04 (ddd, J=3, 10, 59 Hz, 2H), 3.52 (bs, 2H); LC/MS [M+H]=538.

Example 78 Synthesis of 1-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)ethane-1,1,2-tricarboxylic acid

1-(((2R 3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)ethane-1,1,2-tricarboxylic acid was prepared according to the procedure described for Example 2. Isolated as a white solid.

¹H NMR (D₂O, 300 MHz) δ 8.49 (bs, 1H), 5.95-5.96 (d, J=1 Hz, 1H), 4.60-4.49 (m, 1H), 4.30-4.34 (m, 2H), 3.61-3.68 (m, 2H), 2.92-2.94 (m, 2H); LC/MS [M−H]=462.

Example 79 Synthesis of (S)-2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(1H-tetrazol-5-yl)acetic acid and (R)-2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(1H-tetrazol-5-yl)acetic acid

Step 1:

To a solution of 2-chloro-2′,3′-O-isopropylideneadenosine (7.66 g, 22.4 mmol) in DMF (70 mL) at 0° C., under argon atmosphere was added imidazole (3.8 g, 56 mmol) and followed by TBDMSCl (4.033 g, 26.89 mmol), which was added in 4 equal portions over 30 minutes. The reaction mixture was stirred for 2 hours at 0° C. and then allowed to warm up to room temperature and stirred for 18 hours. The solvent was removed on the rotary evaporator and the remainder was quenched with H₂O (150 mL) and extracted with EtOAc (3×150 mL). The combined organic layer was washed further with H₂O (100 mL), brine (100 mL), dried over Na₂SO₄ and concentrated to provide 9-((3aR,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-2-chloro-9H-purin-6-amine (10.78 g) as a white solid.

Step 2:

To a solution of 9-((3aR,4R,6R,6aR)-6-(((tert-butyldimethylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)-2-chloro-9H-purin-6-amine (10.2 g, 22.37 mmol) in dry DMF (150 mL) was added Et₃N (6.8 mL, 48.42 mmol) and 4-DMAP (273 mg, 2.24 mmol). The mixture under argon atmosphere was cooled to 0° C. and followed by dropwise addition of a solution of Boc₂O (10.36 g, 47.45 mmol) in dry DMF (5 mL). The reaction mixture was stirred for 1 hour at 0° C. and then at ambient for 18 hours. The solvent was then removed on the rotary evaporator and the remainder was purified by silica gel column chromatography (22-40% EtOAc in hexanes) to provide N6,N6-bis-Boc-5′-O-tert-butyldimethylsilyl-2′-3′-O-isopropylidene-2-chloro-adenosine (10.36 g) as a solid.

Step 3:

N6,N6-bis-Boc-5′-O-tert-butyldimethylsilyl-2′-3′-O-isopropylidene-2-chloro-adenosine (9.66 g, 14.72 mmol) was dissolved in dry THF (80 mL) and to this mixture was added a solution of TBAF (22.1 mL, 22.1 mmol, 1 M in THF) dropwise. The reaction mixture was stirred 18 hours before it was evaporated to dryness. The remainder was purified by silica gel column chromatography (33-70% EtOAc in hexanes) to provide N6,N6-bis-Boc-2′-3′-O-isopropylidene-2-chloro-adenosine (7.18 g) as a solid.

Step 4:

To a solution of N6,N6-bis-Boc-2′-3′-O-isopropylidene-2-chloro-adenosine (2.5 g, 4.61 mmol) in toluene (15 mL) was added ethyl cyanodiazoacetate (770 mg, 5.35 mmol) and the flask was evacuated and back filled with argon. Rh₂(OAc)₄ (41 mg, 0.09 mmol) was added and the flask was again evacuated and back-filled with argon. The resulting mixture was stirred at 90° C. for 1.5 hours and then over-night at ambient temperature. Additional amounts of ethyl cyanoacetate (200 mg, 1.44 mmol) and Rh₂(OAc)₄ (41 mg, 0.09 mmol) were added and the mixture was heated at 90° C. for an additional 4 hours before it was allowed to cool to room temperature. The crude mixture was purified by silica gel column chromatography (10-70% EtOAc in hexanes) to provide ethyl 2-(((3aR,4R,6R,6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-cyanoacetate (1.22 g) as a mixture of diastereomers (1:1 ratio).

Step 5:

To a solution of diastereomeric mixture of ethyl 2-(((3aR,4R,6R,6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-cyanoacetate (211 mg, 0.323 mmol) in dry toluene (2 mL) was added azidotrimethylsilane (90 uL, 0.65 mmol) and bis(tributyltin) oxide (58 mg, 0.097 mmol). The resulting mixture was stirred at 90° C. for 4 hours. Additional azidotrimethylsilane (90 uL, 0.65 mmol) and bis(tributyltin) oxide (20 mg, 0.033 mmol) were added and the mixture was stirred at 90° C. for 24 hours. The resultant material was concentrated under reduced pressure and the remainder was purified by preparative silica gel TLC (1% MeOH in EtOAc) to provide ethyl 2-(((3aR,4R 6R,6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-(1H-tetrazol-5-yl)acetate (80 mg) as a mixture of diastereomers as a solid (1:1 ratio).

Step 6:

To a solution of diastereomeric mixture of ethyl 2-(((3aR,4R, 6R, 6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-(1H-tetrazol-5-yl)acetate (80 mg, 0.115 mmol) in THF (1 mL) at room temperature was added a solution of 1N aq. NaOH (0.29 mL, 0.29 mmol). The resulting mixture was stirred for 30 minutes before it was acidified to pH ˜6 with 1N aq. HCl solution and concentrated under reduced pressure to provide 2-(((3aR,4R,6R,6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-(1H-tetrazol-5-yl)acetic acid as a diastereomeric mixture (1:1 ratio). This crude product was used in the following step without further purification.

Step 7:

To a solution of diastereomeric mixture of 2-(((3aR,4R,6R,6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-(1H-tetrazol-5-yl)acetic acid (0.115 mmol) in CH₂Cl₂ (0.5 mL) was added TFA (0.5 mL) and H₂O (2 drops). The resulting mixture was stirred for 4 hours and then concentrated under reduced pressure. The crude residue was purified by preparative reverse-phase HPLC to provide a mixture of diastereomers (S)-2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(1H-tetrazol-5-yl)acetic acid and (R)-2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(1H-tetrazol-5-yl)acetic acid (25 mg) as a solid (1:1 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (1:1 ratio) for Example 78 8 9.27 (d, 1H, diastereomeric), 6.07-6.13 (m, 1H), 5.66-5.69 (m, 1H), 4.61-4.70 (m, 1H), 4.36-4.55 (m, 1H), 4.36-4.33 (m, 1H), 3.93-4.20 (m, 1H), 3.79-3.87 (m, 1H); LC/MS [M+H]=428.

Example 80 Synthesis of (S)-2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoic acid; (R)-2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoic acid; (S)-2-(((2R,3S,4R,5R)-5-(6-amino-2-methoxy-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoic acid and (R)-2-(((2R,3S,4R,5R)-5-(6-amino-2-methoxy-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoic acid

Step 1:

To a solution of diastereomeric mixture of ethyl 2-(((3aR,4R,6R,6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-cyanoacetate (1.09 g, 1.67 mmol) in DMF (7.5 mL) at 25° C. was added Cs₂CO₃ (816 mg, 2.504 mmol) and BnBr (0.22 mL, 1.837 mmol). The reaction mixture was stirred for 2 hours. Diluted brine solution (25 mL) was added and the material was extracted with EtOAc (3×25 mL). The combined organic layer was washed further with brine (25 mL), dried over Na₂SO₄ and concentrated. The resulting crude was purified by silica gel column chromatography (5-60% EtOAc in hexanes) to provide (S)-ethyl 2-(((3aR,4R,6R,6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-cyano-3-phenylpropanoate and (R)-ethyl 2-(((3aR,49 6R,6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-cyano-3-phenylpropanoate as a mixture of diastereomers (ca. 1:1 ratio).

Step 2:

The above diastereomeric mixture of (S)-ethyl 2-(((3aR,4R, 6R 6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-cyano-3-phenylpropanoate and (R)-ethyl 2-(((3aR,4R,6R,6aR)-6-(N6,N6-bis-Boc-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-2-cyano-3-phenylpropanoate (817 mg, 1.099 mmol) in dry toluene (2 mL) was added azidotrimethylsilane (0.29 mL, 2.2 mmol) and bis(tributyltin) oxide (197 mg, 0.33 mmol). The resulting mixture was stirred at 80° C. for 1 hour. Additional amounts of azidotrimethylsilane (0.29 mL, 2.2 mmol) and bis(tributyltin) oxide (50 mg, 0.08 mmol) were added and the mixture was stirred at 80° C. for 4.5 hours. Additional azidotrimethylsilane (0.29 mL, 2.2 mmol) was added and the mixture was heated further for 2.5 hours. Additional azidotrimethylsilane (3×0.29 mL, 3×2.2 mmol) and were added and the mixture was stirred at 70-80° C. over a period of 44 hours. The material was cooled to ambient temperature and the crude was purified via preparative silica gel TLC (1% MeOH in EtOAc) to provide ethyl 2-(((3aR,4R,6R,6aR)-6-(6-((tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoate (754 mg, solid) as a mixture of diastereomers (ca. 1:1 ratio).

Step 3:

To a solution of diastereomeric mixture of ethyl 2-(((3aR,4R,6R,6aR)-6-(6-((tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-2,2-dimethyltetrahydrofuro[3,4-d][1,3]dioxol-4-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoate (605 mg, 0.77 mmol) in CH₂Cl₂ (4 mL) was added TFA (4 mL) and H₂O (4 drops). The resulting mixture was stirred for 2 hours and then concentrated under reduced pressure. To the residue was added CH₂Cl₂ (10 mL) and the solvent was then concentrated (cycle repeated 5 times) to provide ethyl 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoate as a mixture of diastereomers (ca. 1:1 ratio).

Step 4:

To a solution of above diastereomeric mixture of ethyl 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoate (420 mg, 0.77 mmol) in MeOH (5 mL) at room temperature was added a solution of 2N aq. NaOH (1.9 mL, 3.9 mmol). The resulting mixture was warmed to 62° C. and stirred for 16 hours. Additional 2N aq. NaOH (1.0 mL, 2.0 mmol) was added. The mixture was heated to 70° C. and stirred for 6 hours. The mixture was cooled to ambient temperature and acidified to pH ˜6 with 1N aq. HCl solution and concentrated under reduced pressure. The crude residue was purified by preparative reverse-phase HPLC to provide two fractions. The first fraction eluted contained a diastereomeric mixture of (S)-2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoic acid (Example 80-a) and (R)-2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoic acid (Example 80-b) as a solid (ca. 1:1 ratio). The second fraction eluted contained a diastereomeric mixture of (S)-2-(((2R,3S,4R,5R)-5-(6-amino-2-methoxy-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoic acid (Example 80-d) and (R)-2-(((2R,3S,4R,5R)-5-(6-amino-2-methoxy-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoic acid (Example 80-c) as a solid (ca. 1:1 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 1:1 ratio) for Examples 80-a and 80-b S 8.37 (d, 1H, diastereomeric), 7.00-7.38 (m, 5H), 5.92-6.04 (m, 1H), 4.67-4.81 (m, 1H), 4.35-4.46 (m, 1H), 4.16-4.30 (m, 1H), 3.93-4.20 (m, 2H), 3.86-3.98 (m, 1H), 3.60-3.81 (m, 2H); LC/MS [M+H]=518.

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 1:1 ratio) for Examples 80-c and 80-d S 8.44 (d, 1H, diastereomeric), 7.02-7.25 (m, 5H), 5.97-6.05 (m, 1H), 4.71-4.83 (m, 1H), 4.37-4.51 (m, 1H), 4.21-4.32 (m, 1H), 4.12 (d, 3H, diastereomeric), 3.57-3.96 (m, 4H); LC/MS [M+H]=514.

Example 82 Synthesis of 2-benzyl-2-(((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

Step 1:

2,6-Dichloro-9-(2-C-methyl-2,3,5-tri-O-benzoyl-β-D-ribofuranosyl)purine (3.0 g, 5.65 mmol, 1 eq) in DMF (30 mL) was added K₂CO₃ (0.82 g, 5.93 mmol, 1.05 eq). To the mixture was added cyclopentanethiol (582.55 mg, 5.70 mmol, 610 uL, 1.01 eq) in DMF (40 mL) at 35° C. dropwise over 30 min. The mixture was stirred at 35° C. for 16 hr. LCMS showed the starting material was consumed completely. The reaction mixture was diluted with Ethyl acetate (100 mL) and washed with saturated aqueous NH₄Cl (50 mL), H₂O (50 mL) and brine (50 mL). The organic layer was separated, dried over Na₂SO₄, and filtered. The filtrate was concentrated in vacuo. The crude mixture was purified by combi-flash on silica gel (Petroleum ether: Ethyl acetate=10:1 to 3:2) to give ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methyl benzoate (2.48 g, 73.5% yield) was obtained as a light yellow gum.

Step 2:

A mixture of ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methyl benzoate (2.28 g, 3.82 mmol, 1 eq) in a solution of saturated NH₃ in MeOH (65.06 mg, 3.82 mmol, 25 mL, 1 eq) was stirred at 0° C. for 2 h, and then at 25° C. for 16h. The reaction mixture was concentrated in vacuo an the resulting gum was dissolved in ethyl acetate (30 mL). The solution was washed with saturated aqueous NH₄Cl (30 mL). The aqueous layer was extracted with ethyl acetate (3×10 mL). The organic combined layer was dried over Na₂SO₄, filtered and concentrated to give a light yellow gum. The crude gum was purified by column chromatography on silica gel to provide (2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-ol (1.8 g, 88.7% yield) as a light yellow gum.

Step 3:

To a solution of (2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-ol (1.34 g, 3.45 mmol, 1 eq) and imidazole (563.04 mg, 8.27 mmol, 2.4 eq) in DMF (13.4 mL) was added TBDPSCl (1.14 g, 4.14 mmol, 1.06 mL, 1.2 eq) in DMF (6.7 mL) at 0° C. The reaction mixture was stirred at 20-25° C. for 16h. Additional amount of TBDPSCl (88 uL, 0.1 eq) was added to the reaction mixture and stirred at 20-25° C. for 3 h. The reaction mixture was quenched with H₂O (70 mL). The aqueous phase was extracted with EtOAc (3×70 mL). The combined organic layer was washed with brine (100 mL), dried over Na₂SO₄, filtered and concentrated. The crude was purified by flash column chromatography on silica gel (eluted with petroleum ether/EtOAc with gradient 1:0-4:1) to provide (2R,3R,4S,5R)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluorotetrahydrofuran-3-ol as a white solid.

Step 4:

To a solution of (2R,3R,4S,5R)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluorotetrahydrofuran-3-ol (1.28 g, 2.04 mmol, 1 eq) and 4-DMAP (37.40 mg, 306.10 umol, 0.15 eq) in DMF (12 mL) was added Boc₂O (668.06 mg, 3.06 mmol, 1.5 eq) and TEA (568.08 uL, 4.08 mmol, 2.0 eq) dropwise at 0° C. The reaction mixture was stirred at 20-25° C. for 1 h. The mixture was washed with H₂O (50 mL). The aqueous phase was extracted with EtOAc (3×30 mL). The combined organic layer was washed with brine (100 mL), dried over Na₂SO₄, filtered and concentrated. The crude was purified by column chromatography on silica gel (eluted with petroleum ether/EtOAc with gradient 1:0-4:1) to provide tert-butyl ((2R,3R,4S,5R)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluorotetrahydrofuran-3-yl) carbonate (1.4 g, 94% yield) as a white foam.

Step 5:

To a solution of tert-butyl ((2R,3R,4S,5R)-2-(((tert-butyldiphenylsilyl)oxy) methyl)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluorotetrahydrofuran-3-yl) carbonate (1.4 g, 1.92 mmol, 1 eq) in THF (14 mL) was added TBAF in THF (1 M, 2.89 mL, 1.5 eq) dropwise at 0° C. The reaction mixture was stirred at 0° C. for 1.5 h. before it was quenched with aq. NH₄Cl (30 mL). The aqueous phase was extracted with EtOAc (3×30 mL). The combined organic layer was washed with brine (50 mL), dried over Na₂SO₄, filtered and concentrated. The crude was purified by column chromatography on silica gel (eluted with petroleum ether/EtOAc with gradient 1:0-4:1) to provide tert-butyl ((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-yl) carbonate as a white foam.

Steps 6-9:

2-Benzyl-2-(((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid was prepared from tert-butyl ((2R,3R,4S,5R)-5-(2-chloro-6-(cyclopentylthio)-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-yl) carbonate according to the procedure described for Example 2. The title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.48 (s, 1H), 7.13-7.29 (m, 5H), 6.48-6.55 (dd, J=3.63 Hz, J=3.60 Hz, 1H), 5.10-5.30 (m, 1H), 4.63-4.72 (m, 1H), 4.31-4.36 (m, 1H), 4.16-4.20 (m, 1H), 3.97-4.02 (m, 2H), 3.40 (s, 2H), 2.29-2.37 (m, 2H), 1.73-1.86 (m, 6H); LC/MS [M+H]=581.1.

Example 83 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2,2-difluorobenzo[d][1,3]dioxol-5-yl)methyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 5-(bromomethyl)-2,2-difluorobenzo[d][1,3]dioxole, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.34 (s, 1H), 7.14 (s, 1H), 7.02-7.04 (d, J=8.14 Hz, 1H), 6.90-6.93 (d, J=8.26 Hz, 1H), 6.44-6.49 (dd, J=4.44 Hz, J=4.29 Hz, 1H), 5.11-5.32 (dt=4.3, 52 Hz, 1H), 4.67-4.76 (m, 1H), 4.10-4.19 (m, 2H), 3.95-3.99 (m, 1H), 3.36-3.49 (q, J=13.44 Hz, 2H); LC/MS [M+H]=576.

Example 84 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carbamoylbenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-(bromomethyl)benzamide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.27 (s, 1H), 7.67-7.70 (d, J=8.02 Hz, 2H), 7.39-7.41 (d, J=8.05 Hz, 2H), 6.39-6.45 (dd, J=4.56, 14.31 Hz, 1H), 5.06-5.26 (t, J=3.90, 53 Hz, 1H), 4.61-4.70 (dt, J=4.10, 17 Hz, J=4.11 Hz, 1H), 4.14-4.19 (m, 1H), 3.98-4.09 (m, 2H), 3.47 (s, 2H); LC/MS [M+H]=539.

Example 85 Synthesis of 2-(((2S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid

Step 1:

To a solution of (2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl) tetrahydrofuran-3-(2 g, 2.91 mmol, 1 eq) in dichloroethane (20 mL) was added di(imidazol-1-yl)methanethione (776.77 mg, 4.36 mmol, 1.5 eq). The resulting mixture was warmed to 70° C. and stirred for 5 hours before it was concentrated to dryness. The residue was purified by flash column (SiO₂, eluted with 40% EtOAc in petroleum ether) to provide O-((2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl) tetrahydrofuran-3-yl) 1H-imidazole-1-carbothioate (1.53 g, 66% yield) as a white solid.

Step 2:

To a solution of O-((2R,3R 4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4-methoxyphenyl)diphenylmethoxy)methyl) tetrahydrofuran-3-yl) 1H-imidazole-1-carbothioate (1.63 g, 2.04 mmol, 1 eq.) in toluene (15 mL) was added AIBN (67.05 mg, 408.30 umol, 0.2 eq.) under nitrogen atmosphere. The reaction was heated at 110° C. and (n-Bu)₃SnH (891.31 mg, 3.06 mmol, 810.28 uL, 1.5 eq.) was added to the reaction carefully dropwise. The reaction was stirred at 110° C. for 1 hour before it was quenched with sat. KF aq. (3 mL) and the reaction mixture was concentrated to dryness. The reaction was purified by flash column (SiO₂, eluted with 30% EtOAc in petroleum ether) to provide 9-((2R,3R,5S)-3-((tert-butyldimethylsilyl)oxy)-5-(((4-methoxyphenyl)diphenylmethoxy)methyl) tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-amine (969 mg, 71% yield) as a white solid.

Step 3:

To a solution of 9-((2R,3R,5S)-3-((tert-butyldimethylsilyl)oxy)-5-(((4-methoxyphenyl)diphenylmethoxy)methyl) tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-amine (969 mg, 1.44 mmol, 1 eq.) in DMF (10 mL) was added TEA (583.40 mg, 5.77 mmol, 802.48 uL, 4 eq.), DMAP (35.22 mg, 288.27 umol, 0.2 eq.) and Boc₂O (1.26 g, 5.77 mmol, 1.32 mL, 4 eq.). The resulting mixture was stirred at 25° C. for 0.5 hour before it was diluted with H₂O (50 mL), extracted with EtOAc (20 mL×4). The combined organic layers were washed with brine (30 mL×2), dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by flash column chromatography (SiO₂, eluted with 20% EtOAc in petroleum ether) to provide the bis-N-Boc protected product (1.05 g, 83% yield) as a white solid.

Step 4:

To a solution of the bis-N-Boc protected product from the previous step (1.7 g, 1.95 mmol, 1 eq.) in DCM (20 mL) was added a solution of TFA (216.39 uL, 2.92 mmol, 1.5 eq.) in DCM (20 mL) at 0° C. dropwise. The resulting mixture was stirred at 25° C. for 1 hour. Additional 100 mg of TFA was added to the reaction and stirred further for 6 hours at 25° C. The reaction was quenched with TEA (2 mL) and concentrated to dryness. The residue was purified by flash column chromatography (SiO₂, eluted with 20% EtOAc in petroleum ether) to provide the desired alcohol (662 mg, 51% yield) as a white solid.

Step 5:

To a solution of the alcohol from the previous step (680 mg, 1.13 mmol, 1 eq.) and Rh₂(OAc)₄ (25.04 mg, 113.30 umol, 0.05 eq.) in toluene (8 mL) was added a solution of diethyl 2-diazomalonate (274.20 mg, 1.47 mmol, 1.3 eq.) in toluene (3 mL) dropwise at 95° C. under N₂ atmosphere. The resulting mixture was stirred at 95° C. for 5 hours before it was concentrated to dryness. The residue was purified by flash column chromatography (SiO₂, eluted with 20% EtOAc in petroleum ether) to provide diethyl 2-(((2S,4R,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)malonate (490 mg, 54% yield) as a light yellow gum.

Step 6:

To a solution of diethyl 2-(((2S,4R,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy) tetrahydrofuran-2-yl)methoxy)malonate (300 mg, 395.61 umol, 1 eq.) in DMF (3 mL) was added K₂CO₃ (109.35 mg, 791.21 umol, 2 eq.) at 25° C. After stirring for 30 min, methyl 4-(bromomethyl)benzoate (181.24 mg, 791.21 umol, 2 eq.) was added to the reaction mixture. The resulting mixture was stirred at 25° C. for 4 hours to give a white suspension. LCMS showed the reaction was completed. The reaction was diluted with H₂O (20 mL), extracted with EtOAc (10 mL×3). The combined organic layers were washed with brine (15 mL), dried over Na₂SO₄, filtered and concentrated to dryness. The residue was purified by flash column (SiO₂, eluted with 20% EtOAc in petroleum ether) to provide diethyl 2-(((2S,4R,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl)benzyl)malonate (199.3 mg, 55% yield, 99%) as a colorless gum.

Step 7:

To a solution of diethyl 2-(((2S,4R,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy) tetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl)benzyl)malonate (168 mg) in DCM (2 mL) at room temperature was added a solution of TFA (2.5 mL). The resulting mixture was stirred for 4 h before H₂O (1 mL) was added and the reaction mixture was stirred further for 4 h. The reaction was concentrated and azeotroped with DCM (3×10 mL) to provide the crude 2-(((2S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid which was used in the subsequent step without further purification.

¹H NMR (CD₃OD, 300 MHz) δ 8.54 (s, 1H), 7.76-7.79 (d, J=8.26 Hz, 2H), 7.32-7.34 (d, J=7.99 Hz, 2H), 5.96 (s, 1H), 4.63-4.70 (m, 2H), 4.09-4.12 (d, J=10.27 Hz, 1H), 3.76-3.80 (d, J=10 Hz, 1H), 3.48 (s, 2H), 2.39-2.50 (m, 1H), 1.98-2.03 (m, 1H); LC/MS [M+H]=522.

Example 86 Synthesis of 2-(((2S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(benzyloxy)benzyl)malonic acid

Proceeding as described in Example 85 above but substituting methyl 4-(bromomethyl)benzoate with 1-(bromomethyl)-4-(phenylmethoxy)benzene, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.51 (s, 1H), 7.26-7.37 (m, 5H), 7.12-7.14 (d, J=8.56 Hz, 2H), 6.71-6.74 (d, J=8.53 Hz, 2H), 5.95 (s, 1H), 4.92 (s, 2H), 4.59-4.66 (m, 2H), 4.09-4.13 (d, J=10.42 Hz, 1H), 3.74-3.78 (d, J=11.23 Hz, 1H), 3.37-3.43 (m, 2H), 2.40-2.49 (m, 1H), 1.96-2.03 (m, 1H); LC/MS [M+H]=584.

Example 87 Synthesis of 2-(((2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-(bromomethyl)benzamide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.45 (s, 1H), 7.69-7.72 (d, J=8.11 Hz, 2H), 7.35-7.37 (d, J=8.08 Hz, 2H), 6.21-6.26 (d, J=16.42 Hz, 1H), 4.44-4.56 (dd, J=9.34, 24.5 Hz, 1H), 4.21-4.27 (m, 2H), 3.94-3.97 (d, J=8.71 Hz, 1H), 3.46-3.60 (q, J=13.99 Hz, 2H), 1.08-1.15 (d, J=22.18 Hz, 3H); LC/MS [M+H]=554.

Example 88 Synthesis of 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(2H-tetrazol-5-yl)-3-(thiophen-3-yl)propanoic acid

Proceeding as described in Example 80 above but substituting benzyl bromide with 3-(bromomethyl)thiophene, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 3:2 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 3:2 ratio):

Minor isomer: S 8.42 (bs, 1H), 7.25-7.20 (m, 1H), 7.08 (bs, 1H), 6.81 (d, J=4.92 Hz, 1H), 6.01 (d, J=5.64 Hz, 1H), 4.80 (t, J=5.25 Hz, 1H), 4.43 (t, J=4.25 Hz, 1H), 4.31-4.22 (m, 1H), 4.0-3.88 (m, 1H), 3.82-3.68 (m, 3H); LC/MS [M+H]=524.

Major isomer: S 8.39 (bs, 1H), 7.18-7.14 (m, 1H), 7.08 (bs, 1H), 6.78 (d, J=4.95 Hz, 1H), 5.98 (d, J=5.37 Hz, 1H), 4.73 (t, J=5.15 Hz, 1H), 4.36 (t, J=4.31 Hz, 1H), 4.31-4.22 (m, 1H), 4.0-3.88 (m, 1H), 3.82-3.68 (m, 3H); LC/MS [M+H]=524.

Example 89 Synthesis of 2-benzyl-2-(((2R,3R,4S,5R)-5-(2-chloro-6-(pentylthio)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-ethoxy-3-oxopropanoic acid

Proceeding as described in Example 82 above but substituting cyclopentanethiol with pentane-1-thiol, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.48-8.53 (t, J=8.16 Hz, 1H), 7.14-7.26 (m, 5H), 6.49-6.55 (m, 1H), 5.10-5.32 (m, 1H), 4.65-4.73 (m, 1H), 4.17-4.23 (m, 3H), 3.99-4.05 (m, 2H), 3.31-3.40 (m, 4H), 1.76-1.86 (m, 2H), 1.20-1.54 (m, 7H), 0.93-0.98 (t, J=6.9 Hz, 3H); LC/MS [M+H]=611.

Example 90 Synthesis of 2-benzyl-2-(((2R,3R,4S,5R)-5-(2-chloro-6-(pentylthio)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

Proceeding as described in Example 2 above by treating diethyl 2-benzyl-2-(((2R,3R,4S,5R)-3-((tert-butoxycarbonyl)oxy)-5-(2-chloro-6-(pentylthio)-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate with aq. LiOH in THF at room temperature to provide 2-benzyl-2-(((2R 3R,4S,5R)-5-(2-chloro-6-(pentylthio)-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid, the title compound was isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.51 (s, 1H), 7.14-7.31 (m, 5H), 6.48-6.54 (dd, J=4.20, 4.35 Hz, 1H), 5.10-5.30 (dt, J=3.86, 52 Hz, 1H), 4.64-4.73 (dt, J=4.68, 17.94 Hz, 1H), 4.15-4.21 (m, 1H), 3.99-4.06 (m, 2H), 3.31-3.40 (m, 4H), 1.81-1.95 (m, 2H), 1.36-1.54 (m, 4H), 0.90-0.98 (m, 3H); LC/MS [M+H]=583.

Example 91 Synthesis of 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(2H-tetrazol-5-yl)-3-(4-(trifluoromethyl)phenyl) propanoic acid

Proceeding as described in Example 80 above but substituting benzyl bromide with 4-(trifluoromethyl)benzyl bromide, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 1:1 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 1:1 ratio):

Isomer 1: S 8.39 (bs, 1H), 7.52-7.43 (m, 2H), 7.38-7.25 (m, 2H), 6.03-5.94 (m, 1H), 4.80 (t, J=4.75 Hz, 1H), 4.48-4.38 (m, 1H), 4.32-4.21 (m, 1H), 4.05-3.68 (m, 4H);

Isomer 2: S 8.36 (bs, 1H), 7.52-7.43 (m, 2H), 7.38-7.25 (m, 2H), 6.03-5.94 (m, 1H), 4.71 (t, J=4.61 Hz, 1H), 4.48-4.38 (m, 1H), 4.32-4.21 (m, 1H), 4.05-3.68 (m, 4H); LC/MS [M+H]=586.

Example 92 Synthesis of 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-(pyridin-3-yl)-2-(1H-tetrazol-5-yl)propanoic acid

Proceeding as described in Example 80 above, but substituting benzyl bromide with 3-(bromomethyl)pyridine hydrobromide, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 1:1 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 1:1 ratio): S 8.62-8.69 (m, 2H), 8.31-8.09 (m, 2H), 7.73-7.60 (m, 1H), 5.90-5.83 (m, 1H), 4.45-4.12 (m, 2H), 3.88-3.50 (m, 5H); LC/MS [M+H]=519.

Example 93 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-chloro-3-(trifluoromethoxy)benzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-chloro-3-(trifluoromethoxy)benzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.31 (bs, 1H), 7.28-7.37 (m, 4H) 6.42-6.48 (dd, J=4, 10 Hz, 1H), 5.09-5.26 (dt, J=4.7, 52 Hz, 1H), 4.64-4.70 (dt, J=5, 17 Hz, 1H), 4.18 (bs, 1H), 4.03 (bs, 2H), 3.41 (bs, 2H); LC/MS [M+H]=614.1.

Example 94 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-chloro-2,6-difluorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-chloro-2,6-di-fluorobenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.18 (d, J=11.6 Hz, 1H), 6.88-6.96 (m, 2H), 6.33-6.39 (dd, J=4, 10 Hz, 1H), 5.04-5.23 (dt, J=4.7, 52 Hz, 1H), 4.55-4.65 (dt, J=5, 17 Hz, 1H), 4.25-4.31 (m, 2H), 3.93 (bs, 1H), 3.43-3.59 (q, J=15 Hz, 2H); LC/MS [M+H]=566.1.

Example 95 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(2-fluorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 2-fluorobenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.28 (s, 1H), 7.38-7.43 (m, 1H), 7.18-7.20 (m, 2H), 6.96-7.03 (m, 1H), 6.37-6.44 (dd, J=4, 10 Hz, 1H), 5.04-5.24 (dt, J=4.7, 52 Hz, 1H), 4.61-4.67 (dt, J=5, 17 Hz, 1H), 4.13-4.15 (m, 1H), 3.98 (bs, 2H), 3.48 (s, 2H); LC/MS [M+H]=514.1.

Example 96 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3,4-dichlorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 3,4-di-chlorobenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.32 (s, 1H), 7.43 (bs, 1H), 7.15-7.28 (m, 2H), 6.42-6.48 (dd, J=4, 10 Hz, 1H), 5.09-5.29 (dt, J=4.7, 52 Hz, 1H), 4.65-4.73 (dt, J=5, 17 Hz, 1H), 4.13-4.22 (m, 1H), 3.97-4.05 (m, 2H), 3.50 (s, 2H): LC/MS [M+H]=564.1.

Example 97 Synthesis of 1-benzyl 3-(2-morpholinoethyl) 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-(trifluoromethoxy)benzyl)malonate

Step 1:

Under an atmosphere of argon gas, mono-benzyl malonate (1.78 g, 9.17 mmol) was dissolved in anhydrous DMF (11 mL) and cooled in a 0° C. bath. Potassium carbonate (317 mg, 2.29 mmol, 0.25 eq.) was added and allowed to stir for 5 minutes before addition of 4-(2-chloroethyl) morpholine (3.58 g, 19.3 mmol, 2.1 eq.). The resulting mixture was stirred at room temperature for 10 hours. The reaction was diluted with EtOAc (50 mL) and washed with water (2×40 mL) and brine (60 mL). The organic layer was concentrated and purified by silica gel column chromatography to give benzyl (2-morpholinoethyl) malonate (1.57 g, 55% yield).

Step 2:

Under an atmosphere of argon gas, a solution of benzyl (2-morpholinoethyl) malonate (1.57 g, 5.11 mmol) was dissolved in acetonitrile (16 mL) and triethylamine (1.42 mL, 10.22 mmol, 2.0 eq.) and cooled in a −10° C. bath for 15 minutes. A solution of 4-acetamidobenzesulfonyl azide (1.84 g, 7.67 mmol, 1.5 eq.) in acetonitrile (16 mL) was added at a rate of 4 mL/min. The reaction was monitored by TLC and completed in 4 hours. The reaction was concentrated and purified by silica gel column chromatography to give 1-benzyl 3-(2-morpholinoethyl) 2-diazomalonate (844 mg, 50% yield).

Step 3: (2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-yl tert-butyl carbonate (1.27 g, 2.11 mmol) and 1-benzyl 3-(2-morpholinoethyl) 2-diazomalonate (844 mg, 2.53 mmol, 1.2 eq.) were charged into a 25 mL round-bottom flask and azeotroped twice with toluene. The resulting oil was dissolved in toluene (13 mL) under an atmosphere of argon gas.

Rhodium(II) acetate dimer (140 mg, 0.32 mmol, 0.15 eq.) was added to the solution and the resulting mixture was heated at 75° C. for 3 h as the reaction proceeded to completion. The reaction was concentrated to an oil and purified by silica gel column chromatography to give 1-benzyl 3-(2-morpholinoethyl) 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (441 mg, 19% yield).

Step 4: 1-Benzyl 3-(2-morpholinoethyl) 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (441 mg, 0.485 mmol) was dissolved in DMF (6.6 mL) and cooled in a wet ice bath. To this solution was added oven dried cesium carbonate (316 mg, 0.970 mmol, 2.0 eq.) and the resulting suspension was stirred for 30 minutes. To this slurry was added 1-(bromomethyl)-3-(trifluoromethoxy)benzene (157 μL, 0.970 mmol, 2.0 eq.) dropwise. The reaction was allowed to warm to ambient temperature and was held for 14 hours. The reaction was diluted with ethyl acetate (50 mL) and was washed twice with water (40 mL), brine, dried over magnesium sulfate and concentrated. Column chromatography purification over silica gel to give 1-benzyl 3-(2-morpholinoethyl) 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(3-(trifluoromethoxy)benzyl)malonate (480 mg, 91% yield).

Step 5:

1-Benzyl 3-(2-morpholinoethyl) 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(3-(trifluoromethoxy)benzyl)malonate (480 mg, 0.440 mmol) was dissolved in DCM (6.0 mL) followed by dropwise addition of TFA (500 μL). The reaction was stirred for 16 hours and monitored by LC/MS. Upon completion the reaction solution was concentrated to dryness. The crude was purified by the reversed-phase HPLC gave the desired 1-benzyl 3-(2-morpholinoethyl) 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-(trifluoromethoxy)benzyl)malonate.

LC/MS [M+H]=783.3.

Example 98 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-(2-morpholinoethoxy)-3-oxo-2-(3-(trifluoromethoxy)benzyl)propanoic acid

Step 1:

1-Benzyl 3-(2-morpholinoethyl) 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(3-(trifluoromethoxy)benzyl) malonate (248 mg, 0.32 mmol) was dissolved at room temperature in EtOAc (2.5 mL) under a blanket of Argon gas. Pd/C 10% (25 mg) was charged into the round-bottom flask and purged three times with Argon gas. A hydrogen-filled balloon was attached to the round bottom and purged twice. The reaction was monitored for completion by LC/MS. The reaction flask was purged with Argon gas upon completion. The reaction slurry was filtered through a bed of celite and rinsed twice with EtOAc (5.0 mL). The filtrate was concentrated and purified by reversed-phase HPLC to give the desired 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-(2-morpholinoethoxy)-3-oxo-2-(3-(trifluoromethoxy)benzyl)propanoic acid.

LC/MS [M+H]=693.2.

Example 99 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(2,3,4-trifluorobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 2,3,4-tri-fluorobenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 300 MHz) δ 8.30 (bs, 1H), 7.15 (q, J=6.63 Hz, 1H), 6.80 (q, J=8.61 Hz, 1H), 6.42 (dd, J=4.41, 12.97 Hz, 1H), 5.18 (td, J=3.88, 52.34 Hz, 1H), 4.67 (td, J=4.11, 17.65 Hz, 1H), 4.18-4.11 (m, 1H), 4.10-3.93 (m, 2H), 3.55-3.41 (m, 2H); LC/MS [M+H]=550.

Example 100 Synthesis of 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxy-3-methyltetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a solution of 1,2-O-(1-Methylethylidene-α-D-xylofuranose (40 g, 210.31 mmol, 1 eq.) in DMF (300 mL) was added imidazole (35.80 g, 525.78 mmol, 2.5 eq.) and TBDPSCl (69.37 g, 252.37 mmol, 64.83 mL, 1.2 eq.) at 0° C. under N2 atmosphere. The mixture was stirred at 25° C. for 5 h before it was quenched with H₂O (1000 mL). The aqueous phase was extracted with EtOAc (3×250 mL). The combined organic layer was washed with brine (500 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography to provide the product (3aR,5R,6S,6aR)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol (80 g, 88.7% yield) as a colorless syrup.

Step 2:

To a solution of (3aR,5R,6S,6aR)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol (80 g, 186.66 mmol, 1 eq.) in DCM (1000 mL) was added DMP (118.75 g, 279.99 mmol, 86.68 mL, 1.5 eq.) at 0° C. The mixture was stirred at 25° C. for 16 h to give a white suspension. The mixture was quenched with saturated aq. Na₂S₂O₃ (500 mL). The aqueous phase was extracted with DCM (2×500 mL). The combined organic layer was washed with brine (500 mL), dried over Na₂SO₄, filtered and concentrated to dryness. The crude product was purified by silica gel column chromatography (0-17% of EtOAc in petroleum ether) to provide (3aR,5R,6aS)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyldihydrofuro[2,3-d][1,3]dioxol-6(3aH)-one (74.1 g, 93% yield) was obtained as a colorless gum.

Step 3:

To a solution of (3aR,5R,6aS)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyldihydrofuro[2,3-d][1,3]dioxol-6(3aH)-one (40.8 g, 95.65 mmol, 1 eq) in THF (400 mL) was added MeMgBr (3 M, 47.82 mL, 1.5 eq.) at 25° C. under N2 atmosphere. The mixture was stirred at 25° C. for 1 h to give a brown suspension before it was quenched with saturated aq. NH₄Cl (500 mL). The aqueous phase was extracted with EA (3×400 mL). The organic layer was washed with brine (500 mL), dried over Na₂SO₄, filtered and concentrated to dryness. The crude product was purified by silica gel column chromatography (0-10% of EtOAc in petroleum ether) to provide (3aR,5R,6R,6aR)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2,6-trimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol (41.77 g, 98.7% yield) as a colorless gum.

Step 4:

To a solution of (3aR,5R,6R,6aR)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2,6-trimethyltetrahydrofuro[2,3-d][1,3]dioxol-6-ol (15 g, 33.89 mmol, 1 eq.) in DCM (150 mL) and water (15 mL, 832.63 mmol, 24.57 eq.) was added TFA (150 mL, 2.03 mol, 59.78 eq.) dropwised at 0° C. The mixture was stirred at 20° C. for 1 h before it was quenched with saturated aq. NaHCO₃ to pH7. The aqueous phase was extracted with DCM (2×230 mL). The combined organic layer was washed with brine (130 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel column chromatography (0-40% of EtOAc in petroleum ether) to provide (3R,4S,5R)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-4-methyltetrahydrofuran-2,3,4-triol as a yellow gum.

Step 5:

To a solution of (3R,4S,5R)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-4-methyltetrahydrofuran-2,3,4-triol (8.23 g, 20.44 mmol, 1 eq.) in pyridine (80 mL) was added 4-DMAP (7.49 g, 61.33 mmol, 3 eq.) and Ac₂O (19.15 mL, 204.44 mmol, 10 eq.) at 20° C. The mixture was stirred at 20° C. for 16 h before it was quenched with H₂O (530 mL). The aqueous phase was extracted with EA (3×230 mL). The combined organic layer was washed with brine (240 mL), dried over Na₂SO₄, filtered and concentrated to dryness. The crude product was purified by silica gel column chromatography (0-50% of EtOAc in petroleum ether) to provide (3R,4R,5R)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-4-methyltetrahydrofuran-2,3,4-triyl triacetate (10.27 g, 95% yield) as a yellow gum.

Step 6:

To a solution of (3R,4R,5R)-5-(((tert-butyldiphenylsilyl)oxy)methyl)-4-methyltetrahydrofuran-2,3,4-triyl triacetate (10.27 g, 19.43 mmol, 1 eq.) in MCCN (110 mL) was added 2-chloro-9H-purin-6-amine (3.62 g, 21.37 mmol, 1.1 eq.), DBU (8.78 mL, 58.28 mmol, 3 eq.) and TMSOTf (17.55 mL, 97.13 mmol, 5 eq.) at 0° C. The mixture was stirred at 0° C. for 0.5 h. Then the mixture was stirred at 65° C. for 1 h before it was quenched with saturated aq. NaHCO₃(300 mL). The aqueous phase was extracted with EtOAc (2×220 mL). The combined organic layer was washed with brine (230 mL), dried over Na₂SO₄, filtered and concentrated to dryness. The crude product was purified by flash silica gel column chromatography (0-100% of EtOAc in petroleum ether) to provide (2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-3-methyltetrahydrofuran-3,4-diyl diacetate (5.6 g, 28% yield) as a yellow solid.

Step 7:

To a solution of (2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-3-methyltetrahydrofuran-3,4-diyl diacetate (5.1 g, 7.99 mmol, 1 eq.) in DMF (50 mL) was added TEA (5.56 mL, 39.96 mmol, 5 eq.), 4-DMAP (292.89 mg, 2.40 mmol, 0.3 eq.) and Boc₂O (8.72 g, 39.96 mmol, 5 eq.) at 20° C. The mixture was stirred at 20° C. for 1 h to give a yellow suspension was quenched with H₂O (250 mL). The aqueous phase was extracted with EA (3×230 mL). The combined organic layer was washed with brine (250 mL), dried over Na₂SO₄, filtered and concentrated to dryness. The crude product was purified by flash silica gel column chromatography (0-33% of EtOAc in petroleum ether) to provide (2R,3R,4R,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-3-methyltetrahydrofuran-3,4-diyl diacetate (5.59 g, 75% yield) as a foam.

Step 8:

To a solution of (2R,3R,4R,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-3-methyltetrahydrofuran-3,4-diyl diacetate (5.59 g, 6.67 mmol, 1 eq.) in THF (60 mL) was added TBAF (1 M, in THF, 10.00 mL, 1.5 eq.) at 0° C. The reaction mixture was stirred at 0° C. for 1 h before it was diluted with H₂O (150 mL). The aqueous phase was extracted with EA (3×130 mL). The combined organic layer was washed with brine (150 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-33% of EtOAc in petroleum ether) to provide (2R,3R,4R,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-2-(hydroxymethyl)-3-methyltetrahydrofuran-3,4-diyl diacetate (3.09 g, 77% yield) as a white foam.

Step 9:

To a solution of (2R,3R,4R,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-2-(hydroxymethyl)-3-methyltetrahydrofuran-3,4-diyl diacetate (680 mg, 1.13 mmol, 1 eq.) in toluene (5 mL) and Rh₂(OAc)₄ (50.09 mg, 113.33 umol, 0.1 eq.) at 20° C. was added a solution of diethyl 2-diazopropanedioate (316.47 mg, 1.70 mmol, 1.5 eq.) in toluene (5 mL). The reaction mixture was heated at 95° C. for 3 hr before it was allowed to cool to room temperature. The reaction was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-25% of EtOAc in petroleum ether) to provide diethyl 2-(((2R,3R 4R,5R)-3,4-diacetoxy-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-3-methyltetrahydrofuran-2-yl)methoxy)malonate (585 mg, 59% yield) as a white foam.

Step 10:

To a solution of diethyl 2-(((2R,3R,4R,5R)-3,4-diacetoxy-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-3-methyltetrahydrofuran-2-yl)methoxy)malonate (535 mg, 705.65 umol, 1 eq.) in DMF (5 mL) was added K₂CO₃ (195.06 mg, 1.41 mmol, 2 eq.) and stirred at 20° C. for 30 min before bromomethylbenzene (167.62 uL, 1.41 mmol, 2 eq.) was added to the mixture and stirred at 20° C. for 15.5 h. The reaction was quenched with H₂O (10 mL). The aqueous phase was extracted with EtOAc (2×10 mL). The combined organic layer was washed with brine (10 mL), dried over Na₂SO₄ filtered and concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-25% of EtOAc in petroleum ether) to provide diethyl 2-benzyl-2-(((2R,3R 4R,5R)-3,4-diacetoxy-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-3-methyltetrahydrofuran-2-yl)methoxy)malonate as colorless gum.

Step 11:

To a solution of diethyl 2-benzyl-2-(((2R,3R,4R,5R)-3,4-diacetoxy-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino-2-chloro-9H-purin-9-yl)-3-methyltetrahydrofuran-2-yl)methoxy)malonate (1.05 g, 1.24 mmol, 1 eq.) in DCM (10 mL) was added TFA (1.5 mL) at 0° C. The mixture was stirred at 25° C. for 16 h before it was quenched with saturated aq. NaHCO₃until it reached pH7. The aqueous phase was extracted with DCM (2×50 mL). The combined organic layer was washed with brine (50 mL), dried over Na₂SO₄, filtered and concentrated to dryness to provide diethyl 2-benzyl-2-(((2R,3R,4R,5R)-3,4-diacetoxy-5-(6-amino-2-chloro-9H-purin-9-yl)-3-methyltetrahydrofuran-2-yl)methoxy)malonate as a foam which was used in the next step directly without further purification.

Step 12:

The mixture of diethyl 2-benzyl-2-(((2R,3R 4R,5R)-3,4-diacetoxy-5-(6-amino-2-chloro-9H-purin-9-yl)-3-methyltetrahydrofuran-2-yl)methoxy)malonate (736 mg, 1.14 mmol, 1 eq.) in saturated NH₃ in MeOH (10 mL) was stirred at 10° C. for 16 h before the mixture was concentrated to dryness. The crude product was purified by flash silica gel column chromatography (0-100% of EtOAc in petroleum ether) to provide diethyl 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxy-3-methyltetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (448 mg, 67% yield) as a white solid.

Step 13:

To a solution of diethyl 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxy-3-methyltetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (150 mg, 265.96 umol, 1 eq.) in EtOH (2 mL) was added LiOH—H₂O (55.80 mg, 1.33 mmol, 5 eq.) in H₂O (0.5 mL) at 10° C. The mixture was stirred at 50° C. for 4 h before it was concentrated to dryness. The residue was dissolved in H₂O (50 ml). The aqueous phase was extracted with EA (2×50 mL). The aqueous phase was adjusted to pH 2-3 with 1 N aq. HCl. The aqueous phase was extracted with EtOAc (3×50 mL). The combined organic layer was washed with brine (50 mL), dried over Na₂SO₄ and filtered. The filtrated was concentrated to dryness and then lyophilization to provide 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxy-3-methyltetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid as a white solid.

¹H NMR (400 MHz, DMSO-d₆) δ 8.38 (s, 1H), 7.77 (br s, 2H), 7.19 (br d, J=6.53 Hz, 2H), 7.03-7.14 (m, 3H), 5.80 (d, J=8.03 Hz, 1H), 4.42 (d, J=8.03 Hz, 1H), 4.00 (m, 1H), 3.60-3.73 (m, 2H), 3.22-3.40 (m, 2H), 1.11 (s, 3H); LC/MS [M+H]=508.0.

Example 101 Synthesis of 2-(((2S,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a mixture of (2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-4-fluorotetrahydrofuran-3-ol (2.6 g, 4.80 mmol, 1 eq) in MCCN (20 mL) was added TCDI (1.28 g, 7.19 mmol, 1.5 eq) at 0° C., the mixture was stirred at 25° C. for 17 hours. Additional of TCDI (1.28 g, 7.19 mmol, 1.5 eq) was added into the above mixture and the mixture was stirred at 25° C. for 5 hours. The mixture was partitioned between EtOAc (100 mL) and water (80 mL), the aqueous phase was extracted with EtOAc (3×40 mL), the combined extracts were washed with brine (100 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a crude product, which was purified by flash silica gel column chromatography (17-33% of EtOAc in petroleum ether) to give O-((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-4-fluorotetrahydrofuran-3-yl) 1H-imidazole-1-carbothioate (2.3 g, 59% yield) as a light yellow gum.

Step 2;

To a mixture of O-((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-2-(((tert-butyldiphenylsilyl)oxy)methyl)-4-fluorotetrahydrofuran-3-yl) 1H-imidazole-1-carbothioate (300 mg, 459.97 umol, 1 eq) in toluene (5 mL) was added AIBN (15.11 mg, 91.99 umol, 0.2 eq) under N2 atmosphere, the mixture was heated to 110° C. and (n-Bu)₃SnH (182.56 uL, 689.96 umol, 1.5 eq) was added into the above mixture dropwise under N2 atmosphere. The mixture was stirred at 110° C. for before the mixture was quenched with saturated KF (20 mL), partitioned between EtOAc (100 mL) and water (10 mL), the organic phase was washed with brine (2×50 mL), dried over anhydrous Na₂SO₄,filtered and concentrated under reduced pressure to give a crude product, which was purified by flash silica gel chromatography (0-40% of EtOAc in petroleum ether) to give 9-((2R,3S,5S)-5-(((tert-butyldiphenylsilyl)oxy) methyl)-3-fluorotetrahydrofuran-2-yl)-2-chloro-9H-purin-6-amine as a white powder.

Step 3:

To a mixture of 9-((2R,3S,5S)-5-(((tert-butyldiphenylsilyl)oxy) methyl)-3-fluorotetrahydrofuran-2-yl)-2-chloro-9H-purin-6-amine (1.20 g, 2.28 mmol, 1 eq) in DMF (10 mL) was added Boc₂O (1.10 g, 5.02 mmol, 2.2 eq), DMAP (83.60 mg, 684.31 umol, 0.3 eq) and TEA (793.73 uL, 5.70 mmol, 2.5 eq). The mixture was stirred at 25° C. under N2 atmosphere for 0.5 hours before the mixture was partitioned between EtOAc (100 mL) and water (50 mL), the aqueous phase was extracted with EtOAc (30 mL×4), and the combined extracts were washed with water (100 mL×2), brine (100 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give the crude bis-N-Boc protected product (1.8 g) as yellow gum which was used for next step directly without purification.

Step 4:

To a mixture of the crude bis-N-Boc protected product (1.68 g, 2.31 mmol, 1 eq) in THF (15 mL) was added TBAF (1 M, 3.47 mL, 1.5 eq) at 0° C., the mixture was stirred at 0° C. for 0.5 hour before the reaction mixture was quenched with cooled water (50 mL), then extracted with EtOAc (3×80 mL), the combined extracts were washed with cooled water (2×80 mL), brine (80 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a crude product as light yellow oil, which was purified by flash silica gel chromatography (0-60% of EtOAc in petroleum ether) to give the desired alcohol as a yellow oil.

Step 5:

To a mixture of the alcohol product from the previous step (850 mg, 1.74 mmol, 1 eq) in toluene (10 mL) was added Rh₂(OAc)₄ (77.00 mg, 174.21 umol, 0.1 eq) and diethyl 2-diazomalonate (421.62 mg, 2.26 mmol, 1.3 eq). The mixture was stirred at 115° C. under N2 atmosphere for 1 hour before the mixture was cooled to room temperature and partitioned between EtOAc (100 mL) and water (20 mL), the organic phase was washed with water (20 mL), brine (20 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a crude product, which was purified by flash silica gel chromatography (0-60% of EtOAc in petroleum ether) to give diethyl 2-(((2S,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate 8 (480 mg, 41% yield) as a light yellow oil.

Steps 6-8:

Proceeding as described in Example 2 above, the title compound was prepared and isolated as a white solid.

LC/MS [M+H]=480.3.

Example 102 Synthesis of 2-(((2S,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3-azido-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Proceeding as described in Example 18 above, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz): δ 8.20 (s, 1H), 7.03-7.20 (m, 2H), 7.05 (dd, J=5.1, 1.6 Hz, 3H), 5.82 (d, J=4.5 Hz, 1H), 4.76-4.78 (m, 1H), 4.48 (br d, J=6.3 Hz, 1H), 4.25 (t, J=5.9 Hz, 1H), 3.88 (d, J=4.3 Hz, 2H), 3.29 (d, J=5.5 Hz, 2H); LC/MS [M+H]=519.4.

Example 103 Synthesis of 2-(((2S,3R,4R,5R)-3-amino-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a colorless solution of diethyl 2-(((2S,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3-azido-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (100 mg, 173.92 umol, 1 eq) in pyridine (2 mL) was added PPh₃ (72.99 mg, 278.27 umol, 1.6 eq). The colorless solution was stirred at 25° C. for 1 hr. To the colorless solution was added NH₄OH (0.40 mL, 2.91 mmol, 16.72 eq). The colorless solution was stirred at 25° C. for 16 h before it was concentrated to give crude diethyl 2-(((2S,3R,4R,5R)-3-amino-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (190 mg) as a colorless gum which was used in next step directly.

Step 2:

To a colorless solution of crude diethyl 2-(((2S,3R,4R,5R)-3-amino-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (190 mg, 173.05 umol, 1 eq) in THF (3 mL) was added LiOH (1 M, 1.73 mL, 10 eq). The colorless solution was stirred at 25° C. for 16 h before it was diluted with water (2 mL), and extracted with ethyl acetate (3×2 mL). To the water layer was added 1N aq. HCl to adjust the pH to 6. The water layer was extracted with ethyl acetate (3×2 mL). The water layer was dried by lyophilization to give white solid (190 mg). This crude was suspended in 10 mL of THF and filtered. The filtration was concentrated to give crude product which was purified by reversed-phase HPLC to provide 2-(((2S,3R,4R,5R)-3-amino-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid as a white solid.

¹H NMR (CD₃OD, 400 MHz): δ 8.32 (br s, 1H), 7.15 (br s, 5H), 5.93 (br s, 1H), 4.50-4.60 (m, 2H), 4.02-4.10 (m, 1H), 3.89 (br s, 1H), 3.78 (br d, J=10.0 Hz, 1H), 3.18-3.27 (m, 2H); LC/MS [M+H]=493.4.

Example 104 Synthesis of 2-(((2R,3S,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-azido-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Proceeding as described in Example 18 above, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz): δ 8.43 (s, 1H), 7.20-7.30 (m, 2H), 7.05-7.11 (m, 3H), 6.39 (d, J=6.53 Hz, 1H), 4.60 (t, J=7.53 Hz, 1H), 4.45-4.50 (m, 1H), 4.06-4.14 (m, 2H), 3.89-4.01 (m, 1H), 3.33-3.49 (m, 2H); LC/MS [M+H]=518.9.

Example 105 Synthesis of 2-(((2R,3S,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3-fluoro-4-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a solution of (2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-((tert-butyldimethylsilyl)oxy)-2-(((4methoxyphenyl)diphenylmethoxy)methyl) tetrahydrofuran-3-ol (10 g, 14.53 mmol, 1 eq.) in DCM (100 mL) was added 4-DMAP (5.32 g, 43.59 mmol, 3 eq.) and Tf₂O (2.88 mL, 17.43 mmol, 1.2 eq.) at 0° C. The mixture was stirred at 0° C. for 1 hr before it was quenched with H₂O (500 mL). The aqueous phase was extracted with DCM (2×340 mL). The combined organic layer was dried over Na₂SO₄, filtered and concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-60% of EtOAc in petroleum ether) to provide the triflate product (9.62 g, 80.7% yield) was obtained as a white foam.

Step 2:

To a solution of the triflate product from the previous step (6.6 g, 8.05 mmol, 1 eq.) in DMF (70 mL) was added AcONa (6.60 g, 80.45 mmol, 10 eq.) at 25° C. The mixture was stirred at 25° C. for 16 hr before the mixture was diluted with H₂O (250 mL). The aqueous phase was extracted with EA (2×200 mL). The combined organic layer was washed with brine (250 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-50% of EtOAc in petroleum ether) to provide the acetate product (4.35 g, 74% yield) was as a foam.

Step 3:

To a mixture of the acetate product from the previous step (4.35 g, 5.96 mmol, 1 eq.) in DMF (40 mL) was added TEA (3.01 g, 29.78 mmol, 4.15 mL, 5 eq.), 4-DMAP (218.30 mg, 1.79 mmol, 0.3 eq.) and tert-butoxycarbonyl tert-butyl carbonate (6.50 g, 29.78 mmol, 5 eq.) at 0° C. The mixture was stirred at 25° C. for 1 hr it was diluted H₂O (250 mL). The aqueous phase was extracted with EtOAc (3×240 mL). The combined organic layer was washed with brine (250 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-20% of EtOAc in petroleum ether) to provide the bis-N-Boc product (4.4 g, 64% yield) as a foam.

Step 4:

To a solution of the bis-N-Boc product from the previous step (4.4 g, 4.73 mmol, 1 eq.) in MeOH (150 mL) was added TEA (150 mL) and H₂O (50 mL) at 20° C. The mixture was stirred at 20° C. for 1 hr before H₂O (120 mL) was added to the reaction mixture. The aqueous phase was extracted with EtOAc (3×120 mL). The combined organic layer was washed with brine (130 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-20% of EtOAc in petroleum ether) to provide the alcohol product (3.59 g, 84% yield) as a white foam.

Step 5:

To a solution of DAST (1.73 g, 10.74 mmol, 1.42 mL, 6 eq.) in DCM (20 mL) was added pyridine (1.5 mL, 18.58 mmol, 10.39 eq.) and the alcohol product from the previous step (1.59 g, 1.79 mmol, 1 eq.) in DCM (20 mL) at 0° C. The mixture was stirred at 20° C. for 3 hr before the mixture was diluted with H₂O (130 mL). The aqueous phase was extracted with DCM (3×120 mL). The combined organic layer was washed with brine (130 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-20% of EtOAc in petroleum ether) to provide the fluoride product (344 mg, 22% yield) was obtained as a yellow foam.

Step 6:

To a solution of the fluoride product from the previous step (344 mg, 386.30 umol, 1 eq.) in DCM (3 mL) was added TFA (150 uL) in DCM (3 mL) at 0° C. The mixture was stirred at 0° C. for 5 min before TEA was added to the reaction mixture to adjust the pH to 7. Then the mixture was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-25% of EtOAc in petroleum ether) to provide the primary alcohol product (226 mg, 68% yield) as a yellow solid.

Steps 7-11:

Proceeding as described in Example 1 above, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz): δ 8.46 (br s, 1H), 7.84 (br s, 2H), 7.13-7.25 (m, 5H), 5.94 (br d, J=6.63 Hz, 1H), 5.85 (d, J=8.00 Hz, 1H), 4.94-5.17 (m, 1H), 4.77-4.91 (m, 1H), 4.37-4.48 (m, 1H), 3.66 (br s, 2H), 3.21 (br s, 2H); LC/MS [M+H]=495.9.

Example 106 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-chloro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a solution of (6aR,8R,9R,9aR)-8-(6-amino-2-chloro-9H-purin-9-yl)-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-9-yl trifluoromethanesulfonate (5 g, 7.39 mmol, 1 eq) in DMF (60 mL) was added LiCl (1.57 g, 36.97 mmol, 5 eq). The colorless solution was stirred at 25° C. for 16 h before it was poured into 100 mL of water, and extracted with ethyl acetate (100 mL) twice. The organic was washed with water (200 mL), brine (200 mL), dried by Na₂SO₄ and concentrated. The crude was purified by Combi-flash (20 g silica gel, ethyl acetate in petrol ether from 20-60%) to give 2-chloro-9-((6aR,8R,9S,9aR)-9-chloro-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-8-yl)-9H-purin-6-amine (2.85 g, 69% yield) as a white solid.

Step 2:

To a colorless solution of 2-chloro-9-((6aR,8R,9S,9aR)-9-chloro-2,2,4,4-tetraisopropyltetrahydro-6H-furo[3,2-f][1,3,5,2,4]trioxadisilocin-8-yl)-9H-purin-6-amine (2.80 g, 4.98 mmol, 1 eq) in DMF (35 mL) was added 4-DMAP (121.60 mg, 995.31 umol, 0.2 eq), TEA (3.02 g, 29.86 mmol, 4.16 mL, 6 eq) and (Boc)₂O (5.43 g, 24.88 mmol, 5.72 mL, 5 eq). The yellow solution was stirred at 25° C. for 1 hr. The reaction was diluted with water (60 mL) and extracted with ethyl acetate (60 mL) twice. The combined organics were washed with water (100 mL), brine (100 mL), dried by Na₂SO₄, and filtered. The filtrate was concentrated to give crude (4.2 g) as a yellow gum. The crude was purified by Combi-flash (20 g silica gel which was treated with TEA, ethyl acetate in petrol ether from 10-25%) to give the bis-N-Boc product (3.35 g, 85.95% yield) as a white solid.

Step 3:

To a colorless solution of the bis-N-Boc product from the previous step (3.35 g, 4.39 mmol, 1 eq) in THF (35 mL) was added TBAF (1 M, 17.57 mL, 4 eq) at 0° C. The solution was stirred at 0° C. for 1 hr. The reaction was diluted with ice-water (50 mL) and extracted with ethyl acetate (50 mL) twice. The organic was washed with water (100 mL), brine (100 mL), dried by Na₂SO₄, and filtered. The filtration was concentrated to give crude. The crude was purified by Combi-flash (12 g silica gel which was treated with TEA, ethyl acetate in petrol ether from 30-80%) to give the 3,5-diol product (1.95 g, 81% yield) as a white solid.

Step 4:

To a colorless solution of the 3,5-diol product from the previous step (1.94 g, 3.73 mmol, 1 eq) in toluene (40 mL) was added Rh₂(OAc)₄ (247.17 mg, 559.23 umol, 0.15 eq). The green suspension was heated to 50° C. under N2. To the suspension was added diethyl 2-diazomalonate (902.27 mg, 4.85 mmol, 1.3 eq). The green suspension was stirred at 50° C. for 36 h. The reaction was diluted with water (20 mL), and extracted with ethyl acetate (20 mL) twice. The organic was washed with brine (40 mL), dried by Na₂SO₄, and filtered. The filtrate was concentrated to give crude (3.2 g) as a purple gum. The crude was purified by Combi-flash (20 g silica gel which was treated with TEA, ethyl acetate in petrol ether from 20-70%) to give diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-chloro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate (1060 mg, 40% yield) as a yellow gum.

Step 5:

To a yellow solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-chloro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate (1.25 g, 1.84 mmol, 1 eq) in DMF (15 mL) was added K₂CO₃ (509.22 mg, 3.68 mmol, 2 eq). The yellow suspension was stirred at 25° C. for 0.5 hr. To the yellow suspension was added bromomethylbenzene (378.11 mg, 2.21 mmol, 262.57 uL, 1.2 eq). The yellow suspension was stirred at 25° C. for 16 hr. Additional amount of K₂CO₃ (500 mg) was added and the yellow suspension was stirred at 25° C. for additional 6 hr. The yellow suspension was diluted with water (20 mL) and extracted with ethyl acetate (20 mL) twice. The organic was washed with water (40 mL), brine (40 mL), dried by Na₂SO₄, and filtered. The filtrate was concentrated to give crude (2.6 g) as a yellow gum. The crude was purified by Combi-flash (20 g silica gel, ethyl acetate in petrol ether from 15-40%) to give diethyl 2-benzyl-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-chloro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate (855 mg, 59% yield) as a white solid.

Step 6:

To a colorless solution of diethyl 2-benzyl-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-chloro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonate (400 mg, 520.40 umol, 1 eq) in DCM (6.7 mL) was added TFA (1.54 g, 13.51 mmol, 1 mL, 25.95 eq) at 0° C. The yellow solution was stirred at 25° C. for 5 hr. The reaction was diluted with saturated aq. NaHCO₃ (20 mL) and extracted with ethyl acetate (20 mL) for three times. The organic was washed with brine (50 mL), dried by Na₂SO₄, and filtered. The filtrate was concentrated to give crude (330 mg) as a yellow gum. The crude was purified by Combi-flash (4 g silica gel, ethyl acetate in petrol ether from 30-80%) to give 290 mg of colorless gum. The gum was dried by lyophilization to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-chloro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (260 mg, 81.6% yield) as a white solid.

Step 7:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-chloro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonate (110 mg, 193.52 umol, 1 eq) in EtOH (4 mL), was added LiOH (1 M, 1.94 mL, 10 eq). The reaction mixture was stirred at 25° C. for 8 h. To the solution was added 2N HCl(aq.) to adjust the pH to 5, and the mixture was extracted with EtOAc(4×5 mL). The combined organics were dried over hydrous Na₂SO₄, and filtered. The filtration was concentrated. The crude was purified by preparative reversed-phase HPLC, and dried by lyophilization to give 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-chloro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid (22.0 mg, 22% yield) as a white powder.

¹H NMR (DMSO-d₆, 400 MHz): 6.41 (s, 1H), 7.85 (br s, 2H), 7.16-7.22 (m, 2H), 7.10-7.15 (m, 3H), 6.41 (d, J=6.3 Hz, 1H), 6.16 (br s, 1H), 4.75-4.83 (m, 1H), 4.54 (br t, J=7.5 Hz, 1H), 3.97 (m, 1H), 3.91 (m, 2H), 3.24 (s, 2H); LC/MS [M+H]=512.4.

Example 107 Synthesis of 2-(((2R,3R,4S,5R)-5-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a solution of compound (2R,3S,4R,5R)-5-((benzoyloxy)methyl)-3-fluorotetrahydrofuran-2,4-diyl dibenzoate (4 g, 8.61 mmol, 1 eq) in DCM (12 mL) was added HBr (6.56 g, 28.36 mmol, 4.4 mL, 35% purity in AcOH, 3.29 eq). The solution was stirred at 25° C. for 16 hr. The pH of the reaction mixture was adjusted to 8 with saturated NaHCO₃(aq.), then extracted with EtOAc (50 mL×3). The combined organics were washed with brine (50 mL×2). The solution was dried with Na₂SO₄ and filtered. The filtration was concentrated to give the bromide product (3.59 g, crude) as a yellow oil.

Step 2:

To a solution of compound ((2R,3R,4S,5R)-3-(benzoyloxy)-5-bromo-4-fluorotetrahydrofuran-2-yl)methyl benzoate (1.91 g, 10.18 mmol, 1.2 eq.) in anhydrous MCCN (100 mL) at 0° C. was added NaH (407.12 mg, 10.18 mmol, 60% purity, 1.2 eq.). The reaction mixture was stirred further for 20 min. A solution of compound 2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidine (3.59 g, 8.48 mmol, 1 eq.) in MCCN (20 mL) was added dropwise over 10 min. The mixture was stirred at 25° C. for 1.5 h before it was quenched with water (30 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered and concentrated to give a residue as yellow foam. The crude was purified by column chromatography on a 40 g silica gel column (eluted with 10-70% of EtOAc in petroleum ether) to provide ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(2,4-dichloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-fluorotetrahydrofuran-2-yl)methyl benzoate (3.8 g, 84% yield) as a white foam.

Step 3:

To a solution of NH₃ in THF (40 mL) was added the di-chloro product from the previous step (2 g, 3.77 mmol, 1 eq). The reaction mixture was stirred at 25° C. for 16 hours in autoclave. The reaction mixture was concentrated to give a mixture of (2R,3R,4S,5R)-5-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-((benzoyloxy)methyl)-4-fluorotetrahydrofuran-3-yl benzoate and (2R,3R,4S,5R)-5-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-((benzoyloxy)methyl)-4-fluorotetrahydrofuran-3-yl benzoate (2.2 g) as a white solid. This crude mixture was used in the next step directly without further purification.

Step 4:

To a solution of a mixture of (2R,3R,4S,5R)-5-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-((benzoyloxy)methyl)-4-fluorotetrahydrofuran-3-yl benzoate and (2R,3R,4S,5R)-5-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-2-((benzoyloxy)methyl)-4-fluorotetrahydrofuran-3-yl benzoate (6.60 g, 16.22 mmol, 1 eq.) in MeOH (100 mL), was added CH₃₀Na/MeOH (1.5 M, 108.16 mL, 10 eq.) under N2 atmosphere. The reaction mixture was stirred at 25° C. for 1 hr. The pH of the reaction mixture was adjusted to 5-6 with 2N HCl (aq.), extracted with EtOAc (150 mL×4). The organic phase was collected and washed with brine (30 mL×3), dried with anhydrous Na₂SO₄, filtered and concentrated to give a residue (5.5 g) as a yellow oil. The crude was purified by combi-flash on a 40 g silica gel (eluted with 25-70% of EtOAc in petroleum ether) to give (2R,3R,4S,5R)-5-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-ol (2.45 g, yield: 46%) as a white solid.

Steps 5-11:

Proceeding as described in Example 1 above, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 13.49 (br s, 2H), 7.58 (br s, 2H), 7.24-7.30 (m, 1H), 7.13-7.24 (m, 5H), 6.59 (d, J=3.76 Hz, 1H), 6.43 (dd, J=16.69, 4.14 Hz, 1H), 5.95 (br s, 1H), 5.00-5.19 (m, 1H), 4.42 (br d, J=18.07 Hz, 1H), 3.95 (m, 1H), 3.78 (br s, 2H), 3.21 (br s, 2H); LC/MS [M+H]=494.9.

Example 108 Synthesis of 2-(((2R,3R,4S,5R)-5-(4-amino-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid

Proceeding as described in Example 2 above but substituting diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate with diethyl 2-(((2R,3R,4S,5R)-5-(4-N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-7H-pyrrolo[2,3-d]pyrimidin-7-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz): δ 12.30-14.28 (br s, 2H), 7.77 (d, J=8.4 Hz, 2H), 7.57 (br s, 2H), 7.35 (d, J=8.4 Hz, 2H), 7.21-7.30 (m, 1H), 6.59 (d, J=4.0 Hz, 1H), 6.44 (dd, J=16.4, 4.0 Hz, 1H), 5.97 (br s, 1H), 5.00-5.23 (m, 1H), 4.42 (m, 1H), 3.92-4.01 (m, 1H), 3.84 (br d, J=4.4 Hz, 2H), 3.24 (s, 2H); LC/MS [M+H]=538.9.

Example 109 Synthesis of 2-(((2R,3R,4S,5R)-5-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Step 1:

To a mixture 3,5-di-O-benzoyl-2-deoxy-2-fluoro-α-D-arabinofuranosyl bromide (5 g, 10.77 mmol, 1 eq) and 4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidine (2.24 g, 11.84 mmol, 1.1 eq) in anhydrous MCCN (80 mL) was added DBU (4.92 g, 32.30 mmol, 4.87 mL, 3 eq) at 0° C. The mixture was stirred at 0° C. for 5 minutes and then TMSOTf (10.77 g, 48.45 mmol, 8.75 mL, 4.5 eq) was added dropwise. The mixture was stirred at 0° C. for 30 mins, and then stirred at 70° C. for 17 hours before the mixture was cooled to room temperature and diluted with EtAOc (200 mL), and the organic layer was washed with saturated NaHCO₃ (100 mL), brine (100 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a crude product, which was purified by flash silica gel chromatography (0-17% of ethyl acetate in petroleum ether) to give ((2R,3R,4S,5R)-3-(benzoyloxy)-5-(4,6-dichloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4-fluorotetrahydrofuran-2-yl)methyl benzoate (3 g, 51.7% yield) as a colorless oil.

Step 2:

To the product from the previous step (2.5 g, 4.71 mmol, 1 eq) was added saturated NH₃ in MeOH (188.21 umol, 30 mL). The mixture was stirred at 25° C. for 17 hours before the mixture was concentrated under reduced pressure to give a crude product, which was purified by flash silica gel chromatography (0-80% of ethyl acetate in petroleum ether) to give (2R,3R,4S,5R)-5-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-ol as a white powder.

Step 3:

To a mixture of the diol product from the previous step (720 mg, 2.37 mmol, 1 eq) in DMF (10 mL) was added TBDPSCl (582.16 uL, 2.27 mmol) and imidazole (403.53 mg, 5.93 mmol, 2.5 eq). The mixture was stirred at 25° C. for 17 hours before the mixture was partitioned between water (50 mL) and EA (80 mL), the aqueous phase was extracted with EA (3×30 mL), the combined extracts were washed with water (2×80 mL), brine (80 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a crude product, which was purified by flash silica gel chromatography (0-75% of ethyl acetate in petroleum ether) to give (2R,3R,4S,5R)-5-(4-amino-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-2-(((tert-butyldiphenylsilyl) oxy)methyl)-4-fluorotetrahydrofuran-3-ol as white powder.

Step 4:

To a mixture of the product from the previous step (1.32 g, 2.44 mmol, 1 eq) in DMF (10 mL) was added TEA (1.23 g, 12.18 mmol, 1.69 mL, 5 eq), 4-DMAP (89.25 mg, 730.53 umol, 0.3 eq) and tert-butoxycarbonyl tert-butyl carbonate (2.66 g, 12.18 mmol, 2.80 mL, 5 eq). The mixture was stirred at 20° C. for 2 hours before the mixture was partitioned between EA (50 mL) and water (50 mL), the aqueous phase was extracted with EA (15 mL×3), and the combined extracts were washed with water (50 mL×2), brine (50 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (0-14% of ethyl acetate in petroleum ether) to give tri-Boc protected product (1 g, 47% yield) as a colorless oil.

Step 5:

To a mixture of the tri-Boc protected product from the previous step (900 mg, 1.07 mmol, 1 eq) in MeOH (10 mL) was added NH₄F (158.27 mg, 4.27 mmol, 4 eq) at 25° C. The mixture was stirred at 25° C. for 15 hour before the mixture was quenched with cool water (40 mL), extracted with EA (50 mL×3), the combined extracts were washed with water (50 mL×2), brine (50 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (0-25% of ethyl acetate in petroleum ether) to give the primary alcohol product (470 mg, 70% yield) as a foam.

Step 6:

To a mixture of the primary alcohol product from the previous step (360 mg, 596.00 umol, 1 eq) in toluene (5 mL) was added Rh₂(OAc)₄ (26.34 mg, 59.60 umol, 0.1 eq), the mixture was stirred at 95° C., and then diethyl 2-diazomalonate (166.43 mg, 894.00 umol, 1.5 eq) in toluene (2 mL) was added dropwise into the above mixture. The mixture was stirred at 95° C. under N2 atmosphere for 3 hours to give a green mixture. The mixture was cooled to room temperature and concentrated under reduced pressure to give a crude product, which was purified by flash silica gel chromatography (0-20% of ethyl acetate in petroleum ether) to give diethyl 2-(((2R,3R,4S,5R)-5-(4-((N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (330 mg, 68% yield) as colorless oil.

Steps 7-9:

Proceeding as described in Example 2 above but substituting diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate with diethyl 2-(((2R,3R,4S,5R)-5-(4-((N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-6-chloro-1H-pyrazolo[3,4-d]pyrimidin-1-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate, the title compound was prepared and isolated as a white solid.

LC/MS [M+H]=496.1.

Example 110 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-nitrobenzyl)malonic acid

Proceeding as described in Example 2 above but substituting benzyl bromide with 4-nitrobenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz) δ 8.23 (s, 1H), 7.96 (d, J=8.4 Hz, 2H), 7.50 (d, J=8.4 Hz, 2H), 6.43 (dd, J=12.0, 4.80 Hz, 1H), 5.08-5.26 (m, 1H), 4.62-4.74 (m, 1H), 4.13-4.19 (m, 1H), 3.95-4.11 (m, 2H), 3.45-3.57 (m, 2H); LC/MS [M+H]=541.3.

Example 111 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-aminobenzyl)malonic acid

Step 1:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(4-nitrobenzyl)malonate (8.16 g, 9.09 mmol, 1 eq.) in EtOH (100 mL) was added PtO₂ (310.86 mg, 1.37 mmol, 1.51e-1 eq.) under H₂ (15 psi). The mixture was stirred at 25° C. for 4 h before the mixture was filtered through a pad of celite. The filtrate was concentrated to dryness to provide diethyl 2-(4-aminobenzyl)-2-(((2R 3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (7.5 g) as an off-white foam.

Step 2:

To a solution of the amino product from the previous step (1 g, 1.15 mmol, 1 eq.) in DCM (10 mL) was added TFA (1.5 mL, 20.26 mmol, 17.57 eq.) at 0° C. The mixture was stirred at 25° C. for 3 h before the reaction mixture was washed with saturated aq. NaHCO₃ to adjust the pH to 7. The aqueous phase was extracted with EtOAc (3×50 mL). The combined organic layer was washed with brine (50 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-100% of ethyl acetate in petroleum ether) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-aminobenzyl)malonate (512 mg, 78% yield) as a yellow gum.

Step 3:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-aminobenzyl)malonate (512 mg, 903.05 umol, 1 eq.) in EtOH (5 mL) was added LiOH (108.14 mg, 4.52 mmol, 5 eq.) in H₂O (1 mL) at 25° C. The mixture was stirred at 50° C. for 16 h before the mixture was concentrated to dryness. The residue was dissolved in H₂O (50 mL). The aqueous phase was extracted with EtOAc (2×50 mL). The aqueous phase was adjusted to pH to 2-3 with 1 N aq. HCl. The aqueous phase was lyophilizated. The crude product was purified by preparative reversed-phase HPLC to provide 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-aminobenzyl)malonic acid (76.4 mg, 16% yield) as an off-white solid.

¹H NMR (DMSO-d₆, 400 MHz) δ 8.29 (d, J=1.76 Hz, 1H), 7.88 (br s, 2H), 6.92 (d, J=8.53 Hz, 2H), 6.50 (br d, J=7.78 Hz, 2H), 6.34 (dd, J=13.93, 4.64 Hz, 1H), 6.01 (br s, 1H), 5.14-5.31 (m, 1H), 4.51 (br d, J=17.57 Hz, 1H), 3.97-4.02 (m, 1H), 3.80 (m, 2H), 3.07 (s, 2H); LC/MS [M+H]=511.

Example 112 Synthesis of 2-(4-acetamidobenzyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid

Step 1:

To a solution of diethyl 2-(4-aminobenzyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (1.49 g, 1.72 mmol, 1 eq.) in AcOH (15 mL) was added Ac₂O (262.72 mg, 2.57 mmol, 241.03 uL, 1.5 eq.) at 25° C. The mixture was stirred at 70° C. for 2 h before the reaction was quenched with H₂O (100 mL). The aqueous phase was extracted with EA (3×50 mL). The combined organic layer was washed with brine (150 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to provide diethyl 2-(4-acetamidobenzyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (2.35 g) as a yellow oil.

Steps 2-3:

Proceeding as described in Example 2 above from diethyl 2-(4-acetamidobenzyl)-2-(((2R 3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate to 2-(4-acetamidobenzyl)-2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)malonic acid, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 9.83 (s, 1H), 8.27 (d, J=1.76 Hz, 1H), 7.88 (br s, 2H), 7.37 (d, J=8.28 Hz, 2H), 7.12 (d, J=8.53 Hz, 2H), 6.35 (dd, J=13.43, 4.64 Hz, 1H), 5.15-5.35 (m, 1H), 4.51 (m, 1H), 4.01 (q, J=4.85 Hz, 1H), 3.88 (br d, J=4.52 Hz, 2H), 3.20 (s, 2H), 1.99 (s, 3H); LC/MS [M+H]=553.1.

Example 113 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-benzamidobenzyl)malonic acid

Step 1:

To a solution of diethyl 2-(4-aminobenzyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (1 g, 1.15 mmol, 1 eq.) in DCM (20 mL) was added TEA (240.72 uL, 1.73 mmol, 1.5 eq.) and BzCl (200.92 uL, 1.73 mmol, 1.5 eq.) at 0° C. The mixture was stirred at 25° C. for 2 h before the reaction was diluted with H₂O (130 mL). The aqueous phase was extracted with EtOAc (3×130 mL). The combined organic layer was washed with brine (150 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-25% of ethyl acetate in petroleum ether) to provide diethyl 2-(4-benzamidobenzyl)-2-(((2R 3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (780 mg, 65% yield) as a yellow foam.

Steps 2-3:

Proceeding as described in Example 2 above from diethyl 2-(4-benzamidobenzyl)-2-(((2R 3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate to 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-benzamidobenzyl)malonic acid, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz) δ 10.16 (s, 1H), 8.29 (d, J=2.01 Hz, 1H), 7.90-7.94 (m, 2H), 7.87 (br s, 2H), 7.49-7.61 (m, 5H), 7.20 (d, J=8.78 Hz, 2H), 6.36 (dd, J=13.55, 4.52 Hz, 1H), 5.16-5.36 (m, 1H), 4.53 (m, 1H), 3.98-4.06 (m, 1H), 3.91 (br d, J=4.77 Hz, 2H), 3.24 (s, 2H); LC/MS [M+H]=615.1.

Example 114 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(methylsulfonamido)benzyl)malonic acid

Step 1:

To a solution of diethyl 2-(4-aminobenzyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (1.5 g, 1.73 mmol, 1 eq.) in DCM (20 mL) was added TEA (262.51 mg, 2.59 mmol, 361.09 uL, 1.5 eq.), DMAP (21.13 mg, 172.95 umol, 0.1 eq.) and MsCl (401.58 uL, 5.19 mmol, 3 eq.) at 0° C. The mixture was stirred at 25° C. for 2 hr before the mixture was quenched with H₂O (50 mL). The aqueous phase was extracted with DCM (3×50 mL). The combined organic layer was washed with brine (50 mL), dried over Mg₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-40% of ethyl acetate in petroleum ether) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(4-(methylsulfonamido)benzyl)malonate (900 mg, 55% yield) as a white foam.

Steps 2-3:

Proceeding as described in Example 2 above from diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(4-(methylsulfonamido)benzyl)malonate to 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(methylsulfonamido)benzyl)malonic acid, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 9.60 (s, 1H), 8.26 (d, J=1.51 Hz, 1H), 7.87 (br s, 2H), 7.18 (d, J=8.53 Hz, 2H), 7.00 (d, J=8.53 Hz, 2H), 6.35 (dd, J=13.05, 4.77 Hz, 1H), 5.17-5.34 (m, 1H), 4.52 (dt, J=18.63, 4.99 Hz, 1H), 4.02 (m, 1H), 3.89 (br d, J=4.27 Hz, 2H), 3.21 (s, 2H), 2.90 (s, 3H); LC/MS [M+H]=589.

Example 115 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-((ethoxycarbonyl)amino)benzyl)malonic acid

Step 1:

To a solution of diethyl 2-(4-aminobenzyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (1 g, 1.15 mmol, 1 eq.) in Py (20 mL) was added ethyl chloroformate (164.64 uL, 1.73 mmol, 1.5 eq.) at 0° C. The mixture was stirred at 25° C. for 2 hr before the reaction was quenched with H₂O (100 mL). The aqueous phase was extracted with EtOAc (2×50 mL). The combined organic layer was washed with brine (50 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-20% of ethyl acetate in petroleum ether) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(4-((ethoxycarbonyl)amino)benzyl)malonate (665 mg, 61% yield) as a white foam.

Steps 2-3:

Proceeding as described in Example 2 above from diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(4-((ethoxycarbonyl)amino)benzyl)malonate to 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-((ethoxycarbonyl)amino)benzyl)malonic acid, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 9.49 (s, 1H), 8.27 (d, J=1.76 Hz, 1H), 7.87 (br s, 2H), 7.25 (br d, J=8.28 Hz, 2H), 7.11 (d, J=8.53 Hz, 2H), 6.35 (dd, J=13.55, 4.77 Hz, 1H), 5.15-5.34 (m, 1H), 4.46-4.56 (m, 1H), 3.98-4.05 (m, 2H), 4.05-4.12 (q, J=7.19 Hz, 2H), 3.18 (s, 2H), 1.20-1.24 (t, J=7.2 Hz, 3H); LC/MS [M+H]=583.1.

Example 116 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-oxopiperidin-1-yl)benzyl)malonic acid

Step 1:

To a solution of diethyl 2-(4-aminobenzyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (1 g, 1.15 mmol, 1 eq.) in THF (10 mL) was added TEA (240.72 uL, 1.73 mmol, 1.5 eq.) and 5-chlorovaleryl chloride (178.74 uL, 1.38 mmol, 1.2 eq.) at 0° C. The mixture was stirred at 25° C. for 2 hr before the mixture was quenched with H₂O (50 mL). The aqueous phase was extracted with DCM (2×50 mL). The combined organic layer was washed with brine (50 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-25% of ethyl acetate in petroleum ether) to provide diethyl 2-(((2R 3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(4-(5-chloropentanamido)benzyl)malonate (789 mg, 69% yield) as a white foam.

Steps 2:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)-2-(4-(5-chloropentanamido)benzyl)malonate (789 mg, 800.31 umol, 1 eq.) in DMF (10 mL) was added K₂CO₃ (331.82 mg, 2.40 mmol, 3 eq.) at 25° C. The mixture was stirred at 60° C. for 4 hr before the reaction was quenched with H₂O (30 mL). The aqueous phase was extracted with EtOAc (3×20 mL). The combined organic layer was washed with brine (40 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-33% of ethyl acetate in petroleum ether) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-oxopiperidin-1-yl)benzyl)malonate (679 mg, 76% yield) as a yellow gum.

Steps 3-4:

Proceeding as described in Example 2 above from provide diethyl 2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-oxopiperidin-1-yl)benzyl)malonate to 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-oxopiperidin-1-yl)benzyl)malonic acid, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₄, 400 MHz,) δ 14.32-12.09 (br s, 2H), 8.25 (d, J=1.76 Hz, 1H), 7.88 (br s, 2H), 7.20 (d, J=8.53 Hz, 2H), 7.01 (d, J=8.28 Hz, 2H), 6.35 (dd, J=12.55, 4.77 Hz, 1H), 5.17-5.35 (m, 1H), 4.53 (dt, J=18.63, 4.99 Hz, 1H), 4.00 (br d, J=3.76 Hz, 1H), 3.87-3.92 (m, 2H), 3.44-3.49 (m, 2H), 3.26 (s, 2H), 2.31-2.37 (m, 2H), 1.74-1.84 (m, 4H); LC/MS [M+H]=593.1.

Example 117 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-iodobenzyl)malonic acid

Proceeding as described in Example 2 above by substituting benzyl bromide with 4-iodobenzyl bromide, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 8.24 (d, J=2.01 Hz, 1H), 7.88 (br s, 2H), 7.49 (d, J=8.28 Hz, 2H), 7.01 (d, J=8.28 Hz, 2H), 6.34 (dd, J=13.18, 4.64 Hz, 1H), 5.16-5.34 (m, 1H), 4.46-4.56 (m, 1H), 3.97-4.00 (m, 1H), 3.87 (br d, J=4.52 Hz, 2H), 3.20 (s, 2H); LC/MS [M+H]=621.9.

Example 118 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-methoxybenzamido)benzyl)malonic acid

Proceeding as described in Example 113 above by substituting benzoyl chloride with 2-methoxylbenzoyl chloride, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 10.02 (s, 1H), 8.30 (d, J=1.25 Hz, 1H), 7.88 (br s, 2H), 7.61 (dd, J=7.53, 1.51 Hz, 1H), 7.54 (br d, J=8.53 Hz, 2H), 7.43-7.52 (m, 1H), 7.16 (br d, J=7.28 Hz, 3H), 7.05 (t, J=7.40 Hz, 1H), 6.35 (dd, J=13.55, 4.52 Hz, 1H), 5.97-6.13 (m, 1H), 5.14-5.39 (m, 1H), 4.45-4.61 (m, 1H), 4.02 (q, J=4.60 Hz, 1H), 3.88 (s, 3H), 3.79-3.87 (m, 2H), 3.14-3.22 (s, 2H); LC/MS [M+H]=645.1.

Example 118 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(thiophene-2-carboxamido)benzyl)malonic acid

Proceeding as described in Example 113 above by substituting benzoyl chloride with 2-thiophenecarbonyl chloride, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.31 (d, J=1.75 Hz, 1H), 7.84 (dd, J=3.75, 1 Hz, 1H), 7.7 (dd, J=5, 1 Hz, 1H), 7.48 (d, J=8.63 Hz, 2H), 7.28 (d, J=8.63 Hz, 2H), 7.16 (dd, J=5, 3.88 Hz, 1H), 6.43 (dd, J=13.26, 4.38 Hz, 1H), 5.25-5.10 (m, 1H) 4.73-4.65 (m, 1H), 4.17 (m, 1H), 4.09-3.95 (m, 2H), 3.4 (m, 2H); LC/MS [M+H]=621.1.

Example 119 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2′-carboxy-[1,1′-biphenyl]-4-yl)methyl)malonic acid and 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2′-(ethoxycarbonyl)-[1,1′-biphenyl]-4-yl)methyl)malonic acid

Step 1:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-iodobenzyl)malonate (300 mg, 442.58 umol, 1 eq), 2-ethoxycarbonylphenylboronic acid pinacol ester (146.65 mg, 531.09 umol, 1.2 eq) and K₂CO₃ (183.50 mg, 1.33 mmol, 3 eq) in 1, 4-dioxane (3 mL) and H₂O (1 mL) was added Pd(dppf)Cl₂ (324 mg, 442.58 umol, 1 eq) at 20-25° C. under N2 atmosphere. The reaction mixture was then heated at 80° C. and stirred for 3 hr. The reaction mixture was diluted with H₂O (15 mL), extracted with EtOAc (3×10 mL). The combined organic layer was washed with brine (30 mL), dried over Na₂SO₄, filtered and concentrated. The crude was purified by preparative TLC (50% of ethyl acetate in petroleum ether) to provide diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2′-(ethoxycarbonyl)-[1,1′-biphenyl]-4-yl)methyl)malonate (282 mg, 88% yield) as a brownish gum.

Step 2:

To a solution of diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2′-(ethoxycarbonyl)-[1,1′-biphenyl]-4-yl)methyl)malonate (100 mg, 142.83 umol, 1 eq) in THF (1 mL) was added LiOH—H₂O (44.95 mg, 1.07 mmol, 7.5 eq) in H₂O (0.5 mL) at 20-25° C. The reaction mixture was stirred at 50° C. for 19 h before it was cooled to room temperature. The reaction mixture was concentrated and the residue was diluted with H₂O (5 mL). The water was wished with EA (3×5 mL) and then acidified with 1N aq. HCl until pH reached 2-3. The mixture was then extracted with EtOAc (3×5 mL). The combined organic layer was washed with brine (15 mL), dried over anhydrous Na₂SO₄, filtered and concentrated to provide 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2′-carboxy-[1,1′-biphenyl]-4-yl)methyl)malonic acid (52.2 mg, 56% yield) as a white solid

¹H NMR (CD₃OD, 400 MHz,) δ 8.38 (s, 1H), 7.73 (d, J=7.78 Hz, 1H), 7.43-7.54 (m, 1H), 7.35-7.43 (m, 1H), 7.31 (d, J=8.03 Hz, 2H), 7.23 (br d, J=7.53 Hz, 1H), 7.15 (d, J=8.03 Hz, 2H), 6.42 (dd, J=13.18, 4.39 Hz, 1H), 5.06-5.28 (m, 1H), 4.69 (dt, J=17.44, 4.33 Hz, 1H), 4.18 (m, 1H), 3.90-4.07 (m, 2H), 3.39-3.50 (m, 2H); LC/MS [M+H]=616.1.

In a separated reaction, the hydrolysis was carried out for 5 h at room temperature provided the ethyl ester.

LC/MS [M+H]=644.2.

Example 120 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((3′-carboxy-[1,1′-biphenyl]-4-yl)methyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonyl phenylboronic acid pinacol ester with 3-ethoxycarbonylphenylboronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.31 (s, 1H), 8.15 (s, 1H), 7.94 (d, J=7.6 Hz, 1H), 7.67 (d, J=7.6 Hz, 1H), 7.47 (t, J=7.6 Hz, 1H), 7.36-7.44 (m, 4H), 6.43 (dd, J=12, 4.8 Hz, 1H), 5.08-5.28 (m, 1H), 4.68-4.77 (m, 1H), 4.16-4.18 (m, 1H), 4.07-4.11 (m, 1H), 3.98 (m, 1H), 3.40-3.52 (m, 2H); LC/MS [M+H]=616.1.

Example 121 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((4′-carboxy-[1,1′-biphenyl]-4-yl)methyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 4-ethoxycarbonylphenylboronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.33 (s, 1H), 8.03 (m, J=8.28 Hz, 2H), 7.56 (m, J=8.28 Hz, 2H), 7.36-7.45 (m, 4H), 6.42 (dd, J=11.92, 4.64 Hz, 1H), 5.12-5.25 (m, 1H), 4.73 (dt, J=17.94, 4.58 Hz, 1H), 4.17 (m, 1H), 4.03-4.12 (m, 1H), 3.93-4.01 (m, 1H), 3.39-3.51 (m, 2H); LC/MS [M+H]=616.0.

Example 122 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(pyridin-2-yl)benzyl)malonic acid

Proceeding as described in Example 119 above but substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 2-(tributylstannyl)pyridine, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.58 (br d, J=4.52 Hz, 1H), 8.34 (s, 1H), 7.99 (t, J=6.96 Hz, 1H), 7.80 (d, J=8.28 Hz, 1H), 7.71 (br d, J=8.03 Hz, 2H), 7.41-7.56 (m, 3H), 6.42 (dd, J=12.55, 4.02 Hz, 1H), 4.97-5.30 (m, 1H), 4.60-4.75 (m, 1H), 4.17 (q, J=4.35 Hz, 1H), 3.98-4.05 (m, 1H), 3.85-3.98 (m, 1H), 3.34-3.51 (m, 2H); LC/MS [M+H]=573.1.

Example 123 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(pyridin-3-yl)benzyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 3-pyridyl boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 8.84 (br s, 1H), 8.56 (br d, J=4.00 Hz, 1H), 8.27 (d, J=1.63 Hz, 1H), 8.01 (br d, J=8.00 Hz, 1H), 7.87 (br s, 2H), 7.46-7.54 (m, 3H), 7.34 (d, J=8.13 Hz, 2H), 6.36 (dd, J=13.13, 4.75 Hz, 1H), 6.06 (br s, 1H), 5.17-5.36 (m, 1H), 4.49-4.60 (m, 1H), 4.03 (q, J=4.92 Hz, 1H), 3.90 (br d, J=4.38 Hz, 2H), 3.31 (s, 2H); LC/MS [M+H]=573.

Example 124 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(pyridin-4-yl)benzyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 4-pyridyl boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 8.64 (d, J=6.02 Hz, 2H), 8.28 (d, J=1.76 Hz, 1H), 7.88 (br s, 2H), 7.69 (br d, J=4.77 Hz, 2H), 7.60 (d, J=8.03 Hz, 2H), 7.36 (d, J=8.28 Hz, 2H), 6.36 (dd, J=13.18, 4.64 Hz, 1H), 6.07 (br s, 1H), 5.14-5.38 (m, 1H), 4.55 (br d, J=18.82 Hz, 1H), 4.00-4.03 (m, 1H), 3.90 (br s, 2H), 3.32 (br s, 2H); LC/MS [M+H]=573.1.

Example 125 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(pyrimidin-5-yl)benzyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 4-pyrimidinyl boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 9.16 (s, 1H), 9.08 (s, 2H), 8.28 (s, 1H), 7.86 (br s, 2H), 7.63 (br d, J=8.00 Hz, 2H), 7.36 (br d, J=7.88 Hz, 2H), 6.36 (br dd, J=13.07, 4.57 Hz, 1H), 6.05 (br s, 1H), 5.14-5.37 (m, 1H), 4.55 (br d, J=18.39 Hz, 1H), 4.03 (br d, J=4.13 Hz, 1H), 3.86 (br s, 2H), 3.30 (s, 2H); LC/MS [M+H]=574.1.

Example 126 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(5-methyl-1H-pyrazol-4-yl)benzyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 5-methylpyrazole-4-boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.38 (s, 1H) 7.61 (s, 1H) 7.27-7.37 (d, J=8, 2H) 7.18-7.26 (d, J=8, 2H) 6.42 (dd, J=13.18, 4.39 Hz, 1H) 5.08-5.28 (m, 1H) 4.66-4.78 (m, 1H) 4.17 (br d, J=4.77 Hz, 1H) 3.88-4.05 (m, 2H) 3.40 (s, 2H) 2.32 (s, 3H); LC/MS [M+H]=576.

Example 127 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(3,5-dimethyl-1H-pyrazol-4-yl)benzyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 3,5-dimethylpyrazole-4-boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.37 (d, J=1.76 Hz, 1H), 7.34 (d, J=8.03 Hz, 2H), 7.10 (d, J=8.03 Hz, 2H), 6.41 (dd, J=13.68, 4.39 Hz, 1H), 4.91-5.24 (m, 1H), 4.68 (br d, J=18.32 Hz, 1H), 4.17 (br d, J=4.27 Hz, 1H), 3.94-4.04 (m, 2H), 3.34-3.45 (m, 2H), 2.25 (s, 6H); LC/MS [M+H]=589.9.

Example 128 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(3,5-dimethylisoxazol-4-yl)benzyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 3,5-dimethylisoxazole-4-boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.33 (s, 1H), 7.39 (d, J=7.38 Hz, 2H), 7.13 (d, J=7.75 Hz, 2H), 6.41 (dd, J=13.45, 4.57 Hz, 1H), 5.01-5.25 (m, 1H), 4.67 (dt, J=17.85, 4.14 Hz, 1H), 4.16 (br d, J=4.75 Hz, 1H), 3.88-4.09 (m, 2H), 3.39-3.50 (m, 2H), 2.31 (s, 3H), 2.16 (s, 3H); LC/MS [M+H]=591.

Example 129 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(thiophen-2-yl)benzyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 2-thiophene-2-boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 8.30 (d, J=1.75 Hz, 1H), 7.88 (br s, 2H), 7.51 (d, J=5.13 Hz, 1H), 7.40-7.47 (m, 3H), 7.24 (d, J=8.25 Hz, 2H), 7.11 (dd, J=5.00, 3.63 Hz, 1H), 6.36 (dd, J=13.38, 4.63 Hz, 1H), 6.06 (br s, 1H), 5.17-5.37 (m, 1H), 4.55 (br d, J=19.76 Hz, 1H), 3.99-4.06 (m, 1H), 3.88 (br s, 2H), 3.26 (s, 2H); LC/MS [M+H]=578.

Example 130 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((4′-(methylsulfonyl)-[1,1′-biphenyl]-4-yl)methyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 4-(methanesulfonylphenyl)-boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.27 (d, J=1.50 Hz, 1H), 7.92-7.96 (m, 2H), 7.68-7.72 (m, 2H), 7.41 (q, J=8.42 Hz, 4H), 6.41 (dd, J=11.26, 4.88 Hz, 1H), 5.11-5.25 (m, 1H), 4.69-4.84 (m, 1H), 4.07-4.18 (m, 2H), 3.95 (dd, J=10.32, 4.06 Hz, 1H), 3.40-3.55 (m, 2H), 3.31 (s, 3H); LC/MS [M+H]=650.1.

Example 131 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((4′-fluoro-[1,1′-biphenyl]-4-yl)methyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 4-fluorophenyl-boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.34 (d, J=1.00 Hz, 1H), 7.39-7.46 (m, 2H), 7.27-7.37 (m, 4H), 7.07 (t, J=8.78 Hz, 2H), 6.42 (dd, J=11.54, 4.77 Hz, 1H) 5.08-5.30 (m, 1H), 4.73 (dt, J=18.01, 4.93 Hz, 1H), 4.13-4.20 (m, 1H), 4.09 (br d, J=10.29 Hz, 1H), 3.96 (dd, J=10.04, 4.27 Hz, 1H), 3.38-3.51 (m, 2H); LC/MS [M+H]=590.4.

Example 132 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-aminopyrimidin-5-yl)benzyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 2-aminopyrimidine-5-boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.45 (s, 2H), 8.29 (s, 1H), 7.34 (q, J=8.28 Hz, 4H), 6.41 (dd, J=12.55, 4.52 Hz, 1H), 5.04-5.25 (m, 1H), 4.70 (dt, J=17.82, 4.77 Hz, 1H), 4.16 (q, J=4.43 Hz, 1H), 4.03 (br dd, J=9.91, 3.39 Hz, 1H), 3.94 (dd, J=9.66, 4.64 Hz, 1H), 3.33-3.49 (m, 2H); LC/MS [M+H]=589.

Example 132 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-((2′-hydroxy-[1,1′-biphenyl]-4-yl)methyl)malonic acid

Proceeding as described in Example 119 above by substituting 2-ethoxycarbonylphenylboronic acid pinacol ester with 2-hydroxyphenyl-boronic acid pinacol ester, the title compound was prepared and isolated as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 9.47 (s, 1H), 8.29 (s, 1H), 7.89 (br s, 2H), 7.35 (d, J=8.28 Hz, 2H), 7.24 (d, J=8.03 Hz, 2H), 7.08-7.17 (m, 2H), 6.91 (d, J=7.53 Hz, 1H), 6.82-6.87 (m, 1H), 6.36 (dd, J=13.80, 4.77 Hz, 1H), 6.05 (br s, 1H), 5.16-5.37 (m, 1H), 4.51-4.62 (m, 1H), 4.00-4.03 (m, 1H), 3.90 (m, 2H), 3.27 (s, 2H); LC/MS [M+H]=588.1.

Example 133 Synthesis of 2-(((2R,3S,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid

Step 1:

To a solution of (2R,3S,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-2-(((4-methoxyphenyl)diphenylmethoxy)methyl) tetrahydrofuran-3-yl 4-nitrobenzoate (930 mg, 1.01 mmol, 1 eq) in DCM (5 mL) was added TFA (0.6 mL, 8.10 mmol, 8.06 eq) in DCM (5 mL) at 0° C. The reaction mixture was stirred at 0° C. for 0.1 h. The reaction mixture was basified by TEA until pH reached ˜7 and then concentrated. The crude was purified by column chromatography on silica gel (eluted with 0-33% of EtOAc in petroleum ether) to provide (2R,3S,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-2-(hydroxymethyl)tetrahydrofuran-3-yl 4-nitrobenzoate (480 mg, 66% yield) as a brownish foam.

Step 2:

To a solution of the alcohol product from the previous step (480 mg, 735.06 umol, 1 eq) and Rh₂(OAc)₄ (32.49 mg, 73.51 umol, 0.1 eq) in toluene (4.8 mL) was added diethyl 2-diazomalonate (246.31 mg, 1.32 mmol, 1.8 eq) in toluene (2.4 mL) under N2 at 90° C. The reaction mixture was stirred at 90° C. for 4 h. The reaction mixture was concentrated to dryness. The crude was purified by column chromatography on silica gel (eluted with 0-30% of EtOAc in petroleum ether) to provide diethyl 2-(((2R,3S,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-((4-nitrobenzoyl)oxy)tetrahydrofuran-2-yl)methoxy)malonate (250 mg, 39% yield) was obtained as a brownish gum.

Step 3:

To a solution of diethyl 2-(((2R,3S,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-((4-nitrobenzoyl)oxy) tetrahydrofuran-2-yl)methoxy)malonate (250 mg, 308.20 umol, 1 eq) and K₂CO₃ (85.19 mg, 616.40 umol, 2 eq) in DMF (2.5 mL) was stirred at 20-25° C. for 1 h. The reaction mixture was added methyl (4-bromomethyl)benzoate (141.20 mg, 616.40 umol, 2 eq) and stirred at 20-25° C. for 16 hr. The reaction mixture diluted with H₂O (20 mL), extracted with EtAOc (3×10 mL). The combined organic layer was washed with brine (20 mL), dried over Na₂SO₄, filtered and concentrated. The crude was purified by preparative TLC (eluted with 33% of EtOAc in petroleum ether) to provide diethyl 2-(((2R,3S,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl) amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-((4-nitrobenzoyl)oxy)tetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl)benzyl)malonate (170 mg, 56% yield) as a colorless gum.

Step 4:

To a solution of diethyl 2-(((2R,3S,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl) amino)-2-chloro-9H-purin-9-yl)-4-fluoro-3-((4-nitrobenzoyl)oxy) tetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl)benzyl)malonate (170 mg, 177.21 umol, 1 eq) in DCM (2 mL) was added TFA (462.00 mg, 4.05 mmol, 0.3 mL, 22.86 eq) at 0° C. The reaction mixture was stirred at 20-25° C. for 1 h. The reaction mixture was cooled to 0° C. and basified by TEA until pH reached ˜7. The mixture was then concentrated. The crude was purified by preparative TLC (eluted with 50% of EtOAc in petroleum ether) to provide diethyl 2-(((2R,3S,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-((4-nitrobenzoyl)oxy)tetrahydrofuran-2-yl) methoxy)-2-(4-(methoxycarbonyl)benzyl)malonate (125 mg, 93% yield) as a colorless gum.

Step 5:

To a solution of diethyl 2-(((2R,3S,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-((4-nitrobenzoyl)oxy)tetrahydrofuran-2-yl) methoxy)-2-(4-(methoxycarbonyl)benzyl)malonate (125 mg, 164.67 umol, 1 eq) in THF (1.2 mL) was added LiOH—H₂O (69.10 mg, 1.65 mmol, 10 eq) in H₂O (0.7 mL) at 20-25° C. The reaction mixture was stirred at 50° C. for 2 h. The reaction mixture was quenched with H₂O (20 mL) and concentrated. The aq. layer was wished with EtOAc (3×15 mL) and then acidified with 1N aq. HCl until the pH reached 2-3 as white solid formed. The mixture was filtered and the solid was collected and dried to provide 2-(((2R,3S,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid (44.9 mg, 49% yield) as a white solid.

¹H NMR (DMSO-d₆, 400 MHz,) δ 12.82 (br s, 2H), 8.29 (s, 1H), 7.89 (br s, 2H), 7.75 (d, J=8.03 Hz, 2H), 7.28 (br d, J=7.53 Hz, 2H), 6.34 (dd, J=14.31, 4.52 Hz, 1H), 6.06 (br s, 1H), 5.12-5.34 (m, 1H), 4.52 (br d, J=18.07 Hz, 1H), 4.00 (br d, J=4.77 Hz, 1H), 3.76 (br s, 2H), 3.23 (br s, 2H); LC/MS [M+H]=540.

Example 134 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-oxopyrrolidin-1-yl)benzyl)malonic acid

Proceeding as described in Example 116 above by substituting 5-chlorovaleryl chloride with 4-chlorobutyryl chloride, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.26 (s, 1H), 7.32-7.39 (d, J=8.4 Hz, 2H), 7.22-7.31 (d, J=8.4 Hz, 2H), 6.42 (dd, J=11.80, 4.77 Hz, 1H), 5.09-5.27 (m, 1H), 4.70 (br d, J=17.32 Hz, 1H), 4.14 (br d, J=5.02 Hz, 1H), 4.03-4.10 (m, 1H), 3.94 (m, 1H), 3.65-3.83 (m, 2H), 3.39 (br d, J=9.03 Hz, 2H), 2.55 (t, J=8.16 Hz, 2H), 2.07-2.19 (m, 2H); LC/MS [M+H]=579.

Example 135 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(1,1-dioxido-1,2-thiazinan-2-yl)benzyl)malonic acid

Proceeding as described in Example 116 above by substituting 5-chlorovaleryl chloride with 4-chloro-1-butanesulfonyl chloride, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) 6 8.32 (s, 1H), 7.29 (d, J=8.53 Hz, 2H), 7.11 (d, J=8.28 Hz, 2H), 6.41 (dd, J=12.80, 4.52 Hz, 1H), 5.05-5.27 (m, 1H), 4.66 (dt, J=18.01, 4.42 Hz, 1H), 4.15 (q, J==4.35 Hz, 1H), 3.92-4.05 (m, 2H), 3.54-3.65 (m, 2H), 3.47-3.49 (m, 2H), 3.13-3.27 (m, 2H), 2.21-2.29 (m, 2H), 1.78-1.88 (m, 2H); LC/MS [M+H]=629.

Example 136 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-oxotetrahydropyrimidin-1(2H)-yl)benzyl)malonic acid

Step 1:

To a mixture of diethyl 2-(4-aminobenzyl)-2-(((2R,3R,4S,5R)-5-(6-(N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-fluorotetrahydrofuran-2-yl)methoxy)malonate (300 mg, 345 umol, 1 eq) in THF (3 mL) was added 1-chloro-3-isocyanatopropane (53.8 mg, 449 umol, 1.3 eq). The mixture was stirred at 25° C. for 17 hours before it was quenched with H₂O (10 mL). The aqueous phase was extracted with EA (2×20 mL). The combined organic layer was washed with brine (10 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness and purified by flash silica gel chromatography (0-50% of EtOAc in petroleum ether) to give the urea product (260 mg, 76% yield) as a colorless gum.

Step 2:

To a mixture of the urea product isolated from the previous step (260 mg, 263 umol, 1 eq) in DMF (8 mL) was added Cs₂CO₃ (343 mg, 1.05 mmol, 4 eq). The mixture was stirred at 50° C. for 2 hours before it was quenched with H₂O (20 mL). The aqueous phase was extracted with EA (2×20 mL). The combined organic layer was washed with brine (20 mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated to dryness. The crude product was purified by flash silica gel chromatography (0-100% of EtOAc in petroleum ether) to give the cyclized urea product (150 mg, 58% yield) as a colorless gum.

Step 3:

To a mixture of the cyclized urea product from the previous step (150 mg, 158 umol, 1 eq) in DCM (2 mL) was added TFA (882 uL, 11.9 mmol, 75.5 eq). The mixture was stirred at 25° C. for 3 hours before it was quenched with saturated aq. NaHCO₃(5 ml), then extracted with EA (2×30 mL). The combined extracts were washed with brine (2×10 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a crude product. The mixture was purified by preparative TLC (1% MeOH in EtOAc) to give diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-oxotetrahydropyrimidin-1(2H)-yl)benzyl)malonate (60 mg, 58% yield) as a white solid.

Step 4:

To a mixture of diethyl 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-oxotetrahydropyrimidin-1(2H)-yl)benzyl)malonate (90 mg, 138 umol, 1 eq) in THF (2 mL) was added LiOH.H₂O (84 mg, 2.00 mmol, 1 mL, 14.4 eq). The mixture was stirred at 20° C. for 17 hours before the mixture was partitioned between EtOAc (10 mL) and water (10 mL). The aqueous phase was adjusted to pH to 2-3 with 2M aq. HCl solution. The aqueous phase was partitioned between EtOAc (40 mL) and brine (20 mL). The organic layer was dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a crude product. The mixture was lyophilized to give 2-(((2R,3R,4S,5R)-5-(6-amino-2-methyl-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-2-(4-(2-oxotetrahydropyrimidin-1(2H)-yl)benzyl)malonic acid (51.2 mg, 59% yield) as a white powder.

¹H NMR (CD₃OD, 400 MHz,) δ 8.37 (d, J=1.25 Hz, 1H), 7.28 (d, J=8.28 Hz, 2H), 7.05 (d, J=8.53 Hz, 2H), 6.42 (dd, J=12.05, 4.52 Hz, 1H), 5.08-5.31 (m, 1H), 4.58-4.73 (m, 1H), 4.11-4.18 (m, 1H), 3.92-4.06 (m, 2H), 3.53 (t, J=5.65 Hz, 2H), 3.35-3.46 (m, 2H), 3.47-3.33 (m, 2H), 1.90-2.04 (m, 2H); LC/MS [M+H]=594.

Example 137 Synthesis of 2-(((2S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid

Step 1:

To a mixture of diethyl 2-(((2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl)benzyl)malonate (70 mg, 112.18 umol, 1 eq) in MCCN (2 mL) was added TCDI (29.99 mg, 168.27 umol, 1.5 eq) at 20° C. The mixture was stirred at 20° C. for 17 hours. Additional amount of TCDI (29.99 mg, 168.27 umol, 1.5 eq) was added into the above mixture, and the mixture was stirred at 20° C. for 3 hours. The mixture was partitioned between EtOAc (20 mL) and water (10 mL), the aqueous phase was extracted with EtOAc (3×10 mL). The combined extracts were washed with brine (2×20 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give diethyl 2-(((2R,3R,4R,5R)-3-((1H-imidazole-1-carbonothioyl)oxy)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl)benzyl)malonate (75 mg) as yellow oil. The crude product was used in next step without purification.

Step 2:

A mixture of the product isolated from the previous step (150 mg, 204.32 umol, 1 eq) and AIBN (3.36 mg, 20.43 umol, 0.1 eq) in toluene (5 mL), the mixture was heated at 110° C., and then n-Bu₃SnH (89.20 mg, 306.48 umol, 81.09 uL, 1.5 eq) was added to the above mixture. The mixture was stirred at 110° C. for 0.5 hour. The mixture was cooled to room temperature and quenched with saturated aq. KF (8 mL), extracted with EtOAc (3×15 mL), the combined extracts was washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a residue, which was purified by flash silica gel chromatography (0-60% of EtOAc in petroleum ether) to give diethyl 2-(((2S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl) benzyl)malonate (65 mg, 104.77 umol, 51.28% yield, 98% purity) as white gum.

Step 3:

To a mixture of diethyl 2-(((2S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)-2-(4-(methoxycarbonyl) benzyl)malonate (60 mg, 98.68 umol, 1 eq) in THF (2 mL) was added LiOH—H₂O (82.82 mg, 1.97 mmol, 20 eq) in H₂O (2 mL). The mixture was stirred at 25° C. for 20.5 hours. The reaction mixture was adjusted the pH to 2-3 with 2M aq. HCl, then extracted with EtOAc (3×50 mL), dried over anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a crude product which was purified by preparative reversed-phase HPLC and was lyophilized to give 2-(((2S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-4-methyltetrahydrofuran-2-yl)methoxy)-2-(4-carboxybenzyl)malonic acid (11.1 mg, 21% yield) as a white powder.

¹H NMR (DMSO-d₆, 400 MHz,) δ 12.79 (br s, 1H), 8.52 (s, 1H), 7.87 (br s, 2H), 7.73 (d, J=8.0 Hz, 2H), 7.31 (d, J=8.0 Hz, 2H), 6.11 (d, J=16.8 Hz, 1H), 4.43-4.64 (m, 1H), 3.94-4.05 (m, 1H), 3.81-3.88 (m, 1H), 3.35 (s, 2H), 2.32-2.37 (m, 2H), 1.07-1.18 (d, J=22.0 Hz, 3H); LC/MS [M+H]=538.

Example 138 Synthesis of 2-(((2S,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxy-3-methyltetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Proceeding as described in Example 2 above, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.65 (s, 1H), 7.19-7.30 (m, 2H), 7.08-7.16 (m, 3H), 5.97 (s, 1H), 4.29 (d, J=4.52 Hz, 1H), 4.17 (d, J=9.29 Hz, 2H), 3.73-3.84 (m, 1H), 3.39-3.52 (m, 2H), 2.62 (m, 1H), 1.03 (d, J=6.78 Hz, 3H); LC/MS [M+H]=492.

Example 139 Synthesis of 2-(((2S,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-hydroxy-3-methyltetrahydrofuran-2-yl)methoxy)-2-benzylmalonic acid

Proceeding as described in Example 2 above, the title compound was prepared and isolated as a white solid.

¹H NMR (CD₃OD, 400 MHz,) δ 8.34 (s, 1H), 7.19 (br d, J=6.78 Hz, 2H), 6.99-7.09 (m, 3H), 5.78 (d, J=7.28 Hz, 1H), 4.59 (m, 1H), 4.40 (br d, J=8.53 Hz, 1H), 3.94 (br d, J=9.03 Hz, 1H), 3.66-3.72 (m, 1H), 3.45-3.52 (m, 1H), 3.33-3.39 (m, 1H), 2.51-2.58 (m, 1H), 1.14 (d, J=6.78 Hz, 3H); LC/MS [M+H]=492.

Example 140 Synthesis of (S)-2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(1H-tetrazol-5-yl)-3-(4-(trifluoromethoxy)phenyl)propanoic acid

Proceeding as described in Example 80 above, but substituting benzyl bromide with 4-(trifluoromethoxy)benzyl bromide, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 1:1 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 1:1 ratio):

Isomer 1: S 8.37 (bs, 1H), 7.19 (d, J=8.35 Hz, 2H), 6.96 (d, J=8.11 Hz, 2H), 5.99 (d, J=5.79 Hz, 1H), 4.71 (t, J=5.12 Hz, 1H), 4.40 (t, J=4.37 Hz, 1H), 4.31-4.20 (m, 1H), 3.98-3.89 (m, 1H), 3.86-3.64 (m, 3H); LC/MS [M+H]=602.

Isomer 2: S 8.40 (bs, 1H), 7.19 (d, J=8.35 Hz, 2H), 7.07 (d, J=7.99 Hz, 2H), 5.98 (d, J=5.67 Hz, 1H), 4.80 (t, J=5.15 Hz, 1H), 4.45 (t, J=4.17 Hz, 1H), 4.31-4.20 (m, 1H), 3.98-3.89 (m, 1H), 3.86-3.64 (m, 3H); LC/MS [M+H]=602.

Example 141 Synthesis of 4-(2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-carboxy-2-(1H-tetrazol-5-yl)ethyl)benzoic acid

Proceeding as described in Example 79 above, but substituting the tetra-Boc protected alcohol described in Example 77 for N6,N6-bis-Boc-2′-3′-O-isopropylidene-2-chloro-adenosine provided the Rh-catalyzed insertion product. The subsequent deprotecting procedure as described in Example 80 above, but substituting benzyl bromide with methyl-4-(bromomethyl)benzoate, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 3:2 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 3:2 ratio):

Minor isomer: S 8.35 (s, 1H), 7.81 (d, J=8.2 Hz, 2H), 7.20 (d, J=8.14 Hz, 2H), 5.98 (d, J=5.1 Hz, 1H), 4.72 (t, J=5.1 Hz, 1H), 4.41 (t, J=4.29 Hz, 1H), 4.32-4.22 (m, 1H), 4.03-3.67 (m, 4H); LC/MS [M+H]=562.

Major isomer: S 8.38 (s, 1H), 7.70 (d, J=8.23 Hz, 2H), 7.20 (d, J=8.13 Hz, 2H), 6.01 (d, J=5.34 Hz, 1H), 4.79 (t, J=5.15 Hz, 1H), 4.46 (t, J=4.38 Hz, 1H), 4.32-4.22 (m, 1H), 4.03-3.67 (m, 4H); LC/MS [M+H]=562.

Example 142 Synthesis of 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-(4-(benzyloxy)phenyl)-2-(1H-tetrazol-5-yl)propanoic acid

Proceeding similarly to that described in Example 80 above, but substituting benzyl bromide with 4-(benzyloxy)benzyl bromide, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 3:2 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 3:2 ratio):

Minor isomer: 8 8.41 (bs, 1H), 7.41-7.21 (M, 5H), 7.01-6.91 (m, 2H), 6.83-6.62 (m, 2H), 6.01 (d, J=5.25 Hz, 1H), 4.90-4.86 (bs, 2H), 4.76 (t, J=4.89 Hz, 1H), 4.49-4.44 (m, 1H), 4.30-4.25 (m, 1H), 4.0-3.85 (m, 1H), 3.79-3.51 (m, 2H), 3.26-3.15 (in, 1H); LC/MS [M+H]=624.

Major isomer: 8 8.32 (bs, 1H), 7.41-7.21 (M, 5H), 7.01-6.91 (m, 2H), 6.83-6.62 (m, 2H), 5.97 (d, J=5.16 Hz, 1H), 4.90-4.86 (bs, 2H), 4.69 (t, J=4.74 Hz, 1H), 4.49-4.44 (m, 1H), 4.30-4.25 (m, 1H), 4.0-3.85 (m, 1H), 3.79-3.51 (m, 2H), 3.26-3.15 (in, 1H); LC/MS [M+H]=624.

Example 143 Synthesis of 2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(1H-tetrazol-5-yl)-3-(3-(trifluoromethoxy)phenyl)propanoic acid

Proceeding similarly to that described in Example 80 above, but substituting benzyl bromide with 3-(trifluoromethoxy)benzyl bromide, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 3:2 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 3:2 ratio):

Minor isomer: S 8.42 (bs, 1H), 7.35-6.94 (M, 4H), 6.01 (d, J=5.16 Hz, 1H), 4.80-4.75 (m, 1H), 4.44 (t, J=4.24 Hz, 1H), 4.32-4.22 (m, 1H), 3.99-3.87 (m, 1H), 3.83-3.67 (m, 3H); LC/MS [M+H]=601.

Major isomer: S 8.42 (bs, 1H), 7.35-6.94 (M, 4H), 5.98 (d, J=4.92 Hz, 1H), 4.72 (t, J=4.91 Hz, 1H), 4.40 (t, J=4.47 Hz, 1H), 4.32-4.22 (m, 1H), 3.99-3.87 (m, 1H), 3.83-3.67 (m, 3H); LC/MS [M+H]=601.

Example 144 Synthesis of 2-(((2R,3R,4S,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-4-fluoro-3-hydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(1H-tetrazol-5-yl)propanoic acid

Proceeding similarly to that described in Example 80 above, but substituting benzyl bromide with 4-(benzyloxy)benzyl bromide, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 1:1 ratio).

LC/MS [M+H]=520.

Example 145 Synthesis of 4′-(2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-carboxy-2-(2H-tetrazol-5-yl)ethyl)-[1,1′-biphenyl]-2-carboxylic acid

Step 1:

To a solution of ethyl 1H-tetrazole-5-acetate (3 g, 19.21 mmol) in DMF (40 mL) under argon atmosphere was added 2-(trimethylsilyl)ethoxymethyl chloride (4.1 mL, 23.05 mmol) and powdered potassium carbonate (5.31 g, 38.42 mmol). The reaction mixture was stirred overnight. Brine (70 mL) and EtOAc (70 mL) were added and the mixture was shaken and the organic phase isolated. The aqueous phase was extracted with EtOAc (2×70 mL). The combined organic layer was washed consecutively with brine (70 mL) and water (70 mL) and then dried over Na₂SO₄ and concentrated. The residue was purified by silica gel column chromatography (15-48% EtOAc in hexanes) to provide ethyl 2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)acetate (2.379 g) as a light yellow mobile oil.

Step 2:

To a solution of ethyl 2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)acetate (2.379 g, 8.31 mmol) in dry acetonitrile (25 mL) under argon atmosphere was added DBU (1.87 mL, 12.47 mmol) and followed by 4-acetamidobenzenesulfonyl azide (2.395 g, 9.96 mmol) in 3 equal portions over 5 minutes. The reaction mixture was stirred for 3.5 hours and then the solvent was removed (rotary evaporator). The residue was purified by silica gel column chromatography (20% EtOAc in hexanes) to provide ethyl 2-diazo-2-(2-((2-(trimethylsilyl) ethoxy)methyl)-2H-tetrazol-5-yl)acetate (2.316 g) as a light orange thick oil.

Steps 4-6:

Proceeding similarly to that described in Example 77 above, but substituting diethyl 2-diazomalonate with ethyl 2-diazo-2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)acetate and also substituting methyl-4-(bromomethyl)benzoate with methyl 4′-bromomethyl biphenyl-2-carboxylate, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 1:1 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 1:1 ratio):

Isomer 1: S 8.52 (s, 1H), 7.77 (d, J=6.18 Hz, 1H), 7.54-7.07 (m, 7H), 6.05 (d, J=5.76 Hz, 1H), 4.82 (t, J=5.34 Hz, 1H), 4.44-4.38 (m, 1H), 4.31-4.27 (m, 1H), 4.09-3.90 (m, 1H), 3.86-3.60 (m, 3H); LC/MS [M+H]=638.

Isomer 2: S 8.39 (s, 1H), 7.76 (d, J=6.33 Hz, 1H), 7.54-7.07 (m, 7H), 5.99 (d, J=5.61 Hz, 1H), 4.70 (t, J=5.25 Hz, 1H), 4.42-4.38 (m, 1H), 4.28-4.23 (m, 1H), 4.09-3.90 (m, 1H), 3.86-3.60 (m, 3H); LC/MS [M+H]=638.

Example 146 Synthesis of 4-(2-(((2R,3S,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-carboxy-2-(1H-tetrazol-5-yl)ethyl)thiophene-2-carboxylic acid

Proceeding similarly to that described in Example X10 above, but substituting 3-(trifluoromethoxy)benzyl bromide with methyl 4-(bromomethyl)thiophene-2-carboxylate, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 1:1 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 1:1 ratio):

Isomer 1: S 8.46 (s, 1H), 7.53 (bs, 1H), 7.38 (bs, 1H), 6.00 (d, J=4.98 Hz, 1H), 4.74 (t, J=5.12 Hz, 1H), 4.44 (t, J=4.25 Hz, 1H), 4.34-4.24 (m, 1H), 4.03-3.90 (m, 1H), 3.83-3.69 (m, 3H); LC/MS [M+H]=568.

Isomer 2: S 8.49 (s, 1H), 7.58 (bs, 1H), 7.40 (bs, 1H), 6.04 (d, J=5.37 Hz, 1H), 4.81 (t, J=5.12 Hz, 1H), 4.39 (t, J=4.49 Hz, 1H), 4.34-4.24 (m, 1H), 4.03-3.90 (m, 1H), 3.83-3.69 (m, 3H); LC/MS [M+H]=568.

Example 147 Synthesis of 2-(((2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxy-4-methyltetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(2H-tetrazol-5-yl)propanoic acid

Proceeding as described in Example 145 above, but substituting tert-butyl-(tert butoxycarbonyl)(9-((2R,3R,4R,5R)-3,4-bis((tert-butoxycarbonyl)oxy)-5-(hydroxymethyl)-3-methyltetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate for tert-butyl-(tert-butoxycarbonyl)(9-((2R,3R,4R,5R)-3,4-bis((tert-butoxycarbonyl)oxy)-5-(hydroxymethyl) tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate and also substituting methyl 4′-bromomethyl biphenyl-2-carboxylate with benzyl bromide, the title compound was prepared and isolated as a solid mixture of diastereomers (ca. 1:1 ratio).

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 3:2 ratio):

Minor isomer: S 8.47 (bs, 1H), 7.19-6.96 (M, 5H), 6.015 (bs, 1H), 4.33 (d, J=9.07 Hz, 1H), 4.25-4.16 (m, 1H), 4.13-3.67 (m, 4H), 0.98 (bs, 3H); LC/MS [M+H]=532.

Major isomer: S 8.48 (bs, 1H), 7.19-6.96 (M, 5H), 6.04 (bs, 1H), 4.39 (d, J=9.04 Hz, 1H), 4.25-4.16 (m, 1H), 4.13-3.67 (m, 4H), 0.98 (bs, 3H); LC/MS [M+H]=532.

Example 148a and 148b Synthesis of 2-(((2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(2H-tetrazol-5-yl)propanoic acid and 2-(((2R,3R,4R,5R)-5-(6-amino-2-chloro-9H-purin-9-yl)-3,4-dihydroxytetrahydrofuran-2-yl)methoxy)-2-(2-(tert-butyl)-2H-tetrazol-5-yl)-3-phenylpropanoic acid

Step 1:

To a solution of tert-butyl (9-((2R,3R,4S,5R)-4-((N-(tert-butoxycarbonyl)(tert-butoxyl)amino)-3-((tert-butyldimethylsilyl)oxy)-5-(hydroxymethyl)tetrahydrofuran-2-yl)-2-chloro-9H-purin-6-yl)carbamate (622 mg, 0.868 mmol) in dry toluene (3 mL) was added ethyl 2-diazo-2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)acetate (353 mg, 1.13 mmol) and the flask was evacuated and back filled with argon. Rh₂(OAc)₄ (8 mg, 0.017 mmol) was added and the flask was again evacuated and back-filled with argon. The resulting mixture was stirred at 75° C. for 2.5 hours before it was allowed to cool to room temperature. The crude mixture was purified by silica gel column chromatography (5-55% EtOAc in hexanes) to provide the diastereomeric mixture of ethyl 2-(((2R,3S,4R,5R)-5-(6-(N,N′-bis(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)-2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)acetate (757 mg) as an oil.

Step 2:

To a solution of diastereomeric mixture of ethyl 2-(((2R,3S,4R,5R)-5-(6-(N,N′-bis(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)-2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)acetate (757 mg, 0.756 mmol) in dry DMF (5 mL) was added Cs₂CO₃ (1.23 g, 3.78 mmol) and BnBr (207 mg, 1.21 mmol). The reaction mixture was stirred for 3 hours. Diluted brine solution (25 mL) was added and the material was extracted with EtOAc (3×25 mL). The combined organic layer was washed further with brine (25 mL), dried over Na₂SO₄ and concentrated. The resulting crude was purified by silica gel column chromatography (5-60% EtOAc in hexanes) to provide ethyl 2-(((2R,3S,4R,5R)-5-(6-(N,N′-bis-(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)propanoate as a mixture of diastereomers (ca. 1:1 ratio).

Step 3:

Ethyl 2-(((2R,3S,4R,5R)-5-(6-(N,N′-bis-(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-((tert-butyldimethylsilyl)oxy)tetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)propanoate (551 mg, 0.505 mmol) was dissolved in dry THF (8 mL) and to this mixture was added a solution of TBAF (1.16 mL, 1.16 mmol, 1 M in THF) dropwise. The reaction mixture was stirred 1 hour before it was evaporated to dryness. The remainder was purified by silica gel column chromatography (8-60% EtOAc in hexanes) to provide ethyl 2-(((2R,3R,4R,5R)-5-(6-(N,N′-bis-(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-hydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)propanoate (169 mg) as a viscous clear oil.

Steps 4-5:

Proceeding similarly to that described in Example 2 above, the ethyl 2-(((2R,3R,4R,5R)-5-(6-(N,N′-bis-(tert-butoxycarbonyl)-2-chloro-9H-purin-9-yl)-3-((tert-butoxycarbonyl)oxy)-4-hydroxytetrahydrofuran-2-yl)methoxy)-3-phenyl-2-(2-((2-(trimethylsilyl)ethoxy)methyl)-2H-tetrazol-5-yl)propanoate was converted to the title diastereomeric compounds, 148a and 148b and isolated as light brown solids.

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 1:1 ratio) for Example X13a:

Isomer 1: S 8.29 (bs, 1H), 7.22-7.16 (M, 2H), 7.15-7.05 (m, 2H), 7.03-6.98 (m, 1H), 5.98 (bs, 1H), 4.61-4.49 (m, 1H), 4.47 (bs, 1H), 4.27-4.18 (m, 1H), 4.13-4.02 (m, 1H), 3.08 (dd, J=8.29, 8.29 Hz, 1H) 3.74-3.55 (m, 2H); LC/MS [M+H]=518.

Isomer 2: S 8.23 (bs, 1H), 7.22-7.16 (M, 2H), 7.15-7.05 (m, 2H), 7.03-6.98 (m, 1H), 5.95 (bs, 1H), 4.61-4.49 (m, 1H), 4.39 (bs, 1H), 4.27-4.18 (m, 1H), 4.13-4.02 (m, 1H), 3.95 (dd, J=5.06, 9.44 Hz, 1H) 3.74-3.55 (m, 2H); LC/MS [M+H]=518.

¹H NMR (CD₃OD, 300 MHz) for a mixture of diastereomers (ca. 1:1 ratio) for Example X13b: Isomer 1: S 8.25 (bs, 1H), 7.35-7.16 (M, 5H), 5.91 (bs, 1H), 4.46-4.36 (m, 2H), 4.18-4.11 (m, 1H), 4.03-3.91 (m, 1H), 3.85-3.75 (m, 1H), 3.72-3.53 (m, 2H), 1.73 (s, 9H); LC/MS [M+H]=574.

Isomer 2: S 8.18 (bs, 1H), 7.35-7.16 (M, 5H), 5.90 (bs, 1H), 4.46-4.36 (m, 2H), 4.18-4.11 (m, 1H), 4.03-3.91 (m, 1H), 3.85-3.75 (m, 1H), 3.72-3.53 (m, 2H), 1.72 (s, 9H); LC/MS [M+H]=574.

Example 149 Inhibition of the CD73 Enzyme In Vitro

For measurements of soluble CD73 enzyme activity, recombinant CD73 was obtained from R&D Systems, Cat. No. 5795-EN-010. Serial dilutions of test compounds were incubated with recombinant CD73 and AMP in reaction buffer (25 mM Tris HCl pH7.5, 5 mM MgCl2, 50 mM NaCl, 0.25 mM DTT, 0.005% Triton X-100). The final reaction volume was 25 μL and the final concentrations of recombinant CD73 and AMP were 0.5 nM and 50 μM, respectively. Reactions were allowed to proceed for 30 minutes at room temperature before the addition of 100 μL Malachite Green (Cell Signaling Technology, Cat. No. 12776).

After 5 minutes at room temperature, absorbance at 630 nm was determined on a microplate spectrophotometer. The concentration of inorganic phosphate was determined using a phosphate standard curve. The IC₅₀ data is given below in Table 3.

TABLE 3 IC₅₀ Cpd. # Compound (μM) 5

13.089  6

0.130 7

0.121 8

0.363 10

21.63  13

0.403 15

0.308 16

0.257 17

0.699 20

0.109 21

0.193 22

0.521 23

0.206 27

0.218 28

2.648 50

1.911 34

0.097 35

0.531 36

0.057 37

0.718 38

0.686 39

0.160 40

1.280 41

1.790 42

0.119 43

1.040 44

0.273 47

0.343 49

0.839 32

0.199 51

1.870 53

0.448 54

0.220 55

>50     56

0.168 57

0.142 58

0.038 59

0.439 60

0.051 61

2.246 63

1.222 64

0.860 65

0.031 66

0.133 67

0.602 68

0.322 73

8.614 74

1.659 75

0.887 76

0.074 77

0.255 78

0.086 79

0.029 80

0.044 81

2.857 82

3.433 83

6.443 84

7.261 84

8.264 85

0.183 86

0.062 87

0.083 88

0.238 89

0.043 91

0.322 92

0.172 94

0.906 95

0.105 96

0.788 97

3.432 98

0.512 107

42.47   108

0.104 109

0.601 110

0.066 111

0.141 113

0.161 114

1.567 115

0.238 116

0.146 117

63.9   118

>200        119

28.04    120

54.51    122

0.032 123

0.152 124

0.135 125

0.242 126

25.333  127

0.149 128

0.258 129

0.022 130

0.240 131

0.098 133

0.742 134

0.087 135

0.111 136

0.110 139

16.523  140

0.030 141

0.060 142

0.067 143

0.136 144

0.065 145

0.106 146

0.181 149

0.051 150

0.016 151

0.031 152

0.034 153

0.074 154

1.800 155

0.047 156

0.086 157

0.034 158

0.105 159

0.076 160

0.012 162

0.054 165

8.926 166

>10     167

>40     168

1.747 169

0.119 171

0.114 174

<0.020   175

0.018 176

0.056 177

0.048 178

0.016 179

0.009 180

>100    181

0.009 182

>50     188

11.525   189

0.227 190

0.300 191

0.130 192

1.178 193

>50     194

0.273 195

>50     197

2.433 198

0.029 199

0.118 200

0.053 201

0.386 202

>50     203

1.107 204

7.847

In Table 4, the IC₅₀ data is as follows: A is <1 μM. B is 1-3 μM, and C is >3 μM.

TABLE 4 IC₅₀ Compound (μM)

A

A

C

A

Example 81 Biological Activity of Disclosed Compounds In Vitro

The ability of compounds to inhibit endogenous, cell-bound CD73 enzyme activity was demonstrated using SK-MEL-28 cells, which express CD73 on their surface. The day before the experiment, 5000 cells were plated per well in a 96-well plate. Cells were washed twice with 200 μL reaction buffer (20 mM HEPES, pH 7.4, 125 mM NaCl, 1 mM KCl, 2 mM MgCl2, 10 mM glucose) to remove residual inorganic phosphate. After washing, assays contained serial dilutions of test compounds and 100 μM of AMP in a total volume of 200 μL reaction buffer, with a final DMSO concentration ≤0.5%. After 30 minutes at room temperature, supernatant was removed from the cells. A volume of 100 μL Malachite Green (Cell Signaling Technology, Cat. No. 12776) was added to 25 μL of supernatant. After 5 minutes at room temperature, absorbance at 630 nm was determined on a microplate spectrophotometer. The concentration of inorganic phosphate was determined using a phosphate standard curve to determine IC₅₀. Table 5 provides the IC₅₀ data of a representative number of compounds.

TABLE 5 Cpd. IC₅₀ # Compound (μM) 6

0.489 7

0.257 8

0.772 13

2.19 15

1.579 16

1.024 17

1.814 20

0.611 21

0.510 22

1.135 23

0.480 27

0.886 50

6.368 34

0.153 35

0.261 36

0.197 37

0.598 38

1.485 39

0.493 40

2.411 41

4.885 42

0.887 43

9.921 44

1.006 47

0.669 49

2.382 32

2.317 51

4.344 53

2.598 54

0.973 55

>50 56

1.250 57

0.1.250 58

0.853 59

6.984 60

0.512 61

6.974 63

2.006 64

0.588 65

0.196 66

0.426 67

1.309 68

0.924 73

1.243 74

2.557 75

0.990 76

0.305 77

1.331 78

0.234 79

0.162 80

0.177 81

2.109 82

4.122 83

5.196 85

1.368 86

0.626 87

0.549 88

0.443 89

0.070 92

0.202 108

0.246 109

1.912 110

0.361 111

0.182 113

0.161 114

1.567 115

0.872 116

0.267 122

0.147 123

0.387 124

0.636 125

0.836 127

0.408 128

0.827 129

0.086 130

1.045 131

0.285 133

>1 134

1.489 135

0.272 136

1.87 189

0.597 190

0.927 191

0.215 192

7.161 194

1.709 197

2.433 198

0.175 199

0.665 200

0.110 201

1.822 203

4.769

In Table 6, the IC₅₀ data is as follows: A is <1 μM. B is 1-3 μM, and C is >3 μM. ND is not disclosed.

TABLE 6 IC₅₀ Compound (μM)

A

INCORPORATION BY REFERENCE

All publications and patents mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.

EQUIVALENTS

While specific embodiments of the subject invention have been discussed, the above specification is illustrative and not restrictive. Many variations of the invention will become apparent to those skilled in the art upon review of this specification and the claims below.

The full scope of the invention should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations. 

1. A compound of formula (I):

or a pharmaceutically acceptable salt and/or prodrug thereof, wherein X is O, NR⁷ or CR⁷R⁸; Y is O or S; Z is NR¹⁹, O or S; Het is heterocyclyl or heteroaryl; R^(1a) is selected from H, halo, hydroxy, cyano, azido, amino, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, amino-C₁₋₆alkyl, —O—C(O)—O—C₁₋₆alkyl, C₁₋₆acyloxy, C₁₋₆alkoxy, C₂₋₆alkenyl, and C₂₋₆alkynyl; and R^(1b) is selected from H, halo, C₁₋₆alkyl, hydroxy-C₁₋₆alkyl, amino-C₁₋₆alkyl, C₂₋₆alkenyl, and C₂₋₆alkynyl; or R^(1a) and R^(1b), together with the carbon atom to which they are attached, form a C═CH₂ or C═C(H)C₁₋₆alkyl; R^(2a) is selected from H, halo, hydroxy, cyano, azido, amino, C₁₋₆alkyl, hydroxy-C₁₋₆alkyl, amino-C₁₋₆alkyl, C₁₋₆acyloxy, —O—C(O)—O—C₁₋₆alkyl, C₁₋₆alkoxy, C₂₋₆alkenyl, and C₂₋₆alkynyl; R^(2b) is selected from H, halo, C₁₋₆alkyl, hydroxy-C₁₋₆alkyl, amino-C₁₋₆alkyl, C₂₋₆alkenyl, and C₂₋₆alkynyl; or R^(2a) and R^(2b), together with the carbon atom to which they are attached, form a C═CH₂ or C═C(H)C₁₋₆alkyl; R³ is selected from H, alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl, and —(CH₂)—C(O)OR₉; R⁴ is selected from heteroaryl, alkyl, —C(O)OR⁹, —C(O)NR¹¹R¹², —S(O)₂R¹⁰, —P(O(OR¹¹)(OR¹²), and —P(O(OR¹¹)(NR¹³R¹⁵); R⁵ is selected from H, cyano, alkyl, cycloalkylalkyl, heterocyclylalkyl, aralkyl, heteroaralkyl, and —C(O)OR⁹; R₆ is selected from —C(O)OR⁹ and —P(O)(OR¹¹)(OR¹²); each R⁷ and R⁸ is H R⁹ is independently selected from H, alkyl, acyloxyalkyl, alkenyl, alkynyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl, and —(CHR¹³)_(m)—Z—C(O)—R¹⁴; each R¹⁰ is independently selected from alkyl, alkenyl, alkynyl, amino, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, and heteroaralkyl; and each R¹¹ and R¹² is independently selected from H, alkyl, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl and —(CHR¹³)_(m)—Z—C(O)—R¹⁴; or R¹¹ and R¹², together with the atoms to which they are attached, form a 5- to 7-membered heterocyclyl; and each R¹³ is independently H or alkyl; each R¹⁴ is independently selected from alkyl, aminoalkyl, heterocyclyl, and heterocyclylalkyl; R¹⁵ is selected from alkyl, aralkyl, —C(R¹⁶)(R¹⁷)—C(O)O—R¹⁸; each R¹⁶ and R¹⁷ are selected from H, alkyl, amino-alkyl, hydroxy-alkyl, mercapto-alkyl, sulfonyl-alkyl, cycloalkyl, aryl, aralkyl, heterocyclylalkyl, heteroaralkyl, and —(CH₂)C(O)OR⁹; R¹⁸ is selected from H, alkyl, alkoxyalkyl, aminoalkyl, haloalkyl, amido, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl, aralkyl, heteroaryl, heteroaralkyl; R¹⁹ is H or alkyl, preferably H; and m is 1 or 2; provided that either R⁴ is tetrazolyl, or R⁵ is aralkyl or heteroaralkyl, or both. 2.-146. (canceled)
 147. A compound selected from: Cpd. # Compound Name 6

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-benzyl- malonic acid 7

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-(4- carboxybenzyl)-malonic acid 8

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-((6- chloropyridin-3-yl) methyl)malonic acid 10

2-(((2R,3R,4R,5R)-5-(6- amino-9H-purin-9-yl)-4- fluoro-3-hydroxytetra- hydrofuran-2-yl)methoxy)- 2-benzylmalonic acid 12

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4-(2- carboxyethyl)benzyl) malonic acid 13

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- phenoxybenzyl)malonic acid 15

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- (trifluoromethoxy) benzyl)malonic acid 16

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- (trifluoromethyl)benzyl) malonic acid 17

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2- (naphthalen-2-ylmethyl) malonic acid 20

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (trifluoromethyl)benzyl) malonic acid 21

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (methylsulfonyl)benzyl) malonic acid 22

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- (dimethylcarbamoyl) benzyl)malonic acid 23

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2- (thiophen-3-ylmethyl) malonic acid 25

diethyl 2-(((2R,3R,4S,5R)- 5-(6-amino-chloro-9H- purin-9-yl)-4-fluoro-3- (isobutyryloxy)tetrahydro- furan-2-yl)methoxy)-2-(4- (trifluoromethoxy)benzyl) malonate 27

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- aminobenzyl)malonic acid 28

2-benzyl-2- (((2R,3R,4S,5R)-5-(6- (benzylamino)-2- chloro-9H-purin-9-yl)- 4-fluoro-3-hydroxytetra- hydrofuran-2-yl) methoxy)malonic acid 32

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-((6- (trifluoromethyl)pyridin- 3-yl)methyl)malonic acid 34

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- cyanobenzyl)malonic acid 35

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- cyanobenzyl)malonic acid 36

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (trifluoromethoxy)benzyl) malonic acid 37

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-3- ethoxy-3-oxo-2-(4- (trifluoromethoxy)benzyl) propanoic acid 38

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-3- ethoxy-3-oxo-2-(4- (trifluoromethoxy)benzyl) propanoic acid 39

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (hydroxymethyl)benzyl) malonic acid 40

2-(((2R,3R,4S,5R)-5-(6- amino-2-azido-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- cyanobenzyl)malonic acid 41

2-(((2R,3R,4S,5R)-5-(2- azido-6-((tert-butoxy- carbonyl)amino)-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- cyanobenzyl)malonic acid 42

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro- furan-2-yl)methoxy)-2- (3-carboxybenzyl)- malonic acid 43

2-(3-(1H-tetrazol-5-yl) benzyl)-2- (((2R,3R,4S,5R)-5-(6- amino-2-azido-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)malonic acid 44

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- fluorobenzyl)malonic acid 45

diethyl 2-(((2R,3R,4S,5R)- 5-(6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- fluorobenzyl)malonate 46

diethyl 2-(((2R,3R,4S,5R)- 5-(6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- fluorobenzyl)malonate 47

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- fluorobenzyl)malonic acid 48

diethyl 2-(((2R,3R,4S,5R)- 5-(6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (trifluoromethoxy)benzyl) malonate 49

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- methoxybenzyl)malonic acid 50

2-((1H-tetrazol-5-yl) methyl)-2- (((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)malonic acid 51

2-(((2S,3S,4R,5R)-3- amino-5-(6-amino-2- chloro-9H-purin- 9-yl)-4- hydroxytetrahydrofuran- 2-yl)methoxy)-2- benzylmalonic acid 53

2-(3-(1H-tetrazol-5-yl) benzyl)-2- (((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)malonic acid 54

2-(4-(1H-tetrazol-5-yl) benzyl)-2- (((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro- furan-2-yl)methoxy) malonic acid 55

2-(((2R,3S,4R,5R)-4- amino-5-(6-amino-2- chloro-9H-purin-9-yl)-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2- benzylmalonic acid 56

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- hydroxybenzyl)malonic acid 57

2-(((2R,3R,4S,5R)-5- (6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- carboxy-2-fluorobenzyl) malonic acid 58

2-(((2R,3R,4S,5R)-5- (6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-(4- carboxy-3-fluorobenzyl) malonic acid 59

2-(((2R,3R,4S,5R)-5- (6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-((5- (trifluoromethyl)-furan- 2-yl)methyl)malonic acid 60

2-(((2R,3R,4S,5R)-5- (6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-(3- fluoro-4-(trifluoromethyl) benzyl)malonic acid 61

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-((3- phenylisoxazol-5-yl) methyl)malonic acid 62

dimethyl 2- (((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (trifluoromethyl)benzyl) malonate 63

2-(((2R,3R,4S,5R)-5- (6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-((1- benzyl-1H-pyrazol-4-yl) methyl)malonic acid 64

2-((1H-pyrazol-4-yl) methyl)-2- (((2R,3R,4S,5R)- 5-(6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)malonic acid 65

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (benzyloxy)benzyl) malonic acid 66

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- hydroxybenzyl)malonic acid 67

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-((2- carboxythiazol-5-yl) methyl)malonic acid 68

2-(((2R,3R,4S,5R)-5- (6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-((2- carboxythiazol-4-yl) methyl)malonic acid 69

2-([1,1′-biphenyl]-4- ylmethyl)-2- (((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-3-ethoxy- 3-oxopropanoic acid 70

diethyl 2-(((2R,3R,4S,5R)- 5-(6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4-(2- methoxy-2-oxoethyl) benzyl)malonate 71

diethyl 2-(((2R,3R,4S,5R)- 5-(6-amino-chloro-9H- purin-9-yl)-4-fluoro-3- (isobutyryloxy)tetrahydro- furan-2-yl)methoxy)-2- ((5-(methoxycarbonyl) thiophen-3-yl)methyl) malonate 72

diethyl 2-(((2R,3R,4S,5R)- 5-(6-amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran-2- yl)methoxy)-2-((5- (methoxycarbonyl)-1- methyl-1H-pyrazol-3-yl) methyl)malonate 73

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-((5- carboxy-1-methyl-1H- pyrazol-3-yl)methyl) malonic acid 74

2-benzyl-2- (((2R,3R,4S,5R)-5-(2- chloro-6-hydroxy-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)malonic acid 75

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- cyano-3-fluorobenzyl) malonic acid 76

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-((5- carboxythiophen-3-yl) methyl)malonic acid 77

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-((5- carboxythiophen-2-yl) methyl)malonic acid 78

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (carboxymethyl)benzyl) malonic acid 79

2-([1,1′-biphenyl]-4- ylmethyl)-2- (((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)malonic acid 80

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- chloro-2-fluorobenzyl) malonic acid 81

2-([1,1′-biphenyl]-4- ylmethyl)-2- (((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-3- methoxy-3-oxopropanoic acid 82

2-([1,1′-biphenyl]-4- ylmethyl)-2- (((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-3- methoxy-3-oxopropanoic acid 83

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-((3- carboxyisoxazol-5-yl) methyl)malonic acid 84

2-([1,1′-biphenyl]-4- ylmethyl)-2- (((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-3-ethoxy- 3-oxopropanoic acid 85

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(3- chloro-4-methoxybenzyl) malonic acid 86

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- sulfamoylbenzyl)malonic acid 87

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-(4-((2- carboxy-ethyl)carbamoyl) benzyl)malonic acid 88

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydro-furan- 2-yl)methoxy)-2-((2- carboxybenzofuran-5-yl) methyl)malonic acid 89

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (2,2,2-trifluoroethoxy) benzyl)malonic acid 90

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- ((E)-2-carboxyvinyl) benzyl)malonic acid 91

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (methoxycarbonyl) benzyl)malonic acid 92

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (carboxymethoxy) benzyl)malonic acid 93

2-benzyl-2- (((2R,3R,4S,5R)-5-(2- chloro-6-oxo-1H-purin- 9(6H)-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)malonic acid 94

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4-(5- methyl-1,3,4-oxadiazol- 2-yl)benzyl)malonic acid 95

of 2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-((2′- cyano-[1,1′-biphenyl]- 4-yl)-methyl)malonic acid 96

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-((5- chlorobenzo[b]thiophen- 3-yl)methyl)-malonic acid 97

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(benzo [d]thiazol-2-ylmethyl) malonic acid 98

2-(((2R,3R,4S,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-4-fluoro-3- hydroxytetrahydrofuran- 2-yl)methoxy)-2-(4- (methylcarbamoyl)benzyl) malonic acid 99

Diethyl 2- (((2R,3S,4R,5R)- 5-(6-amino-2-chloro-9H- purin-9-yl)-3,4- dihydroxytetrahydrofuran- 2-yl)methoxy)-2-benzyl malonate 100

2-(((2R,3S,4R,5R)-5-(6- amino-2-chloro-9H- purin-9-yl)-3,4- dihydroxytetrahydro- furan-2-yl)methoxy)-2- (4-carboxybenzyl) malonic acid 101

S)-2-(((2R,3S,4R,5R)- 5-(6-amino-2-chloro- 9H-purin-9-yl)-3,4- dihydroxytetrahydro- furan-2-yl)methoxy)- 2-(1H-tetrazol-5-yl) acetic acid 102

(R)-2-(((2R,3S,4R,5R)- 5-(6-amino-2-chloro- 9H-purin-9-yl)-3,4- dihydroxytetrahydro- furan-2-yl)methoxy)-2- (1H-tetrazol-5-yl)acetic acid 103

(S)-2-(((2R,3S,4R,5R)- 5-(6-amino-2-chloro- 9H-purin-9-yl)-3,4- dihydroxytetrahydro- furan-2-yl)methoxy)- 3-phenyl-2-(1H-tetrazol- 5-yl)propanoic acid 104

(R)-2-(((2R,3S,4R,5R)- 5-(6-amino-2-chloro- 9H-purin-9-yl)-3,4- dihydroxytetrahydro- furan-2-yl)methoxy)-3- phenyl-2-(1H-tetrazol- 5-yl)propanoic acid 105

(S)-2-(((2R,3S,4R,5R)- 5-(6-amino-2-methoxy- 9H-purin-9-yl)-3,4- dihydroxytetrahydro- furan-2-yl)methoxy)-3- phenyl-2-(1H-tetrazol- 5-yl)propanoic acid 106

(R)-2-(((2R,3S,4R,5R)- 5-(6-amino-2-methoxy- 9H-purin-9-yl)-3,4- dihydroxytetrahydro- furan-2-yl)methoxy)-3- phenyl-2-(1H-tetrazol- 5-yl)propanoic acid

Cpd. # Compound 108

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or a pharmaceutically acceptable salt thereof.
 148. A compound selected from:

or a pharmaceutically acceptable salt thereof.
 149. A pharmaceutical composition comprising the compound claim 1, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.
 150. A method of inhibiting CD73 in a cell, comprising contacting the cell with a compound of formula (I), or a pharmaceutically acceptable salt thereof, according to claim
 1. 151. The method of claim 150, wherein contacting the cell occurs in a subject in need thereof, thereby treating a disease or disorder selected from cancer, cerebral and cardiac ischemic diseases, fibrosis, immune and inflammatory disorders, inflammatory gut motility disorder, neurological, neurodegenerative and CNS disorders and diseases, depression, Parkinson's disease, and sleep disorders.
 152. The method of claim 151, wherein the cancer is selected from bladder cancer, bone cancer, brain cancer, breast cancer, cardiac cancer, cervical cancer, colon cancer, colorectal cancer, esophageal cancer, fibrosarcoma, gastric cancer, gastrointestinal cancer, head & neck cancer, Kaposi's sarcoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, myeloma, ovarian cancer, pancreatic cancer, penile cancer, prostate cancer, testicular germcell cancer, thymoma and thymic carcinoma. 153.-158. (canceled) 